Vestec multi-gauge manual

So far, no specific biochemical marker of the phenotype has been shown to be a universal feature of amoeboid cells arising from different cell types. Importantly, cancer cell invasion is responsive to surrounding conditions and transitions between the individual modes can occur. The mesenchymal-amoeboid MAT or amoeboid-mesenchymal AMT transitions can be induced by modulating the activity of key signalling hubs, such as the Rho GTPases, or by targeting necessary mechanisms of either invasion mode 13 , The plasticity of invasion is presumably the main reason why clinically usable anti-metastatic treatment strategies are still unavailable Despite the large effort to reveal signalling underlying invasive behaviour of cells, understanding of cancer cell invasion plasticity is still insufficient, mainly due to the scarcity of results obtained from more in vivo -like 3D cell culture conditions.

To date, there are only three published works reporting gene expression profiling of amoeboid cells 16 , 17 , While these data provided the first insight into the transcriptome of amoeboid cells, they were not obtained from three-dimensional 3D cultures, an essential requirement to get the most relevant results.

To gain more insight into molecular level adaptation of cancer cells to the amoeboid state, we performed large scale transcriptomic and proteomic profiling of HT fibrosarcoma cells after MAT in 3D cell culture Fig. In order to discern treatment-specific effects, we used two experimental treatments that are sufficiently effective in inducing MAT and compatible with cell viability in 3D collagen gels. The first treatment was doxycycline-inducible constitutively active RhoA icaRhoA gene expression; RhoA-ROCK pathway is known to play a key role in amoeboid migration 13 , 19 and constitutively active RhoA expression has been shown to induce amoeboid morphology in glioblastoma cells and effective MAT in HT 3 , The second, very different treatment was that with dasatinib, a Src kinase inhibitor, that has been previously also shown to induce MAT 21 , Protein lysates were trypsin-digested, TMT-labelled, fractionated and analysed on Thermo Orbitrap Fusion mass spectrometer.

Overall, our work provides data for an unprecedented comparison of parallel mesenchymal and amoeboid transcriptomes and proteomes obtained from 3D conditions to reveal new details of the transition regulation, the impact of the transition on biological properties of the cells in terms of gene expression and protein abundance, and possibly discover potential new therapeutic opportunities. The cultures were regularly tested for mycoplasma contamination.

Stably transfected cells were prepared by lentiviral transduction using the second-generation packaging system pLVX constructs, Tet-On Advanced Gene expression system, Clontech.

All populations of stably transfected cells were further enriched for the respective encoded fluorescence with a cell sorter. Cells bearing inducible constructs were transiently induced with doxycycline before the sorting. All plasmids used in the study were constructed in the lab with standard molecular cloning procedures; details as well as the plasmids themselves are available upon request.

Cell morphology was assessed by measuring the ratio of the maximum length and maximum width manually using Fiji software At least cells were analysed for each condition. The quality and size distribution of sequencing libraries was analysed with Agilent BioAnalyzer Raw reads were trimmed of adapter sequences with Cutadapt 25 version 1.

The samples and related files are summarized in Table 1. Protein samples were trypsin-digested, TMT-labeled, fractionated and analysed on Orbitrap Fusion mass spectrometer Thermo Fisher Scientific as described previously 28 , 29 , TMT label was added according to manufacturer protocol. All samples were pooled and vacuum dried. Peptides were fractionated as follows. Most intense precursors were isolated by quadrupole with 1. Raw data were processed in Proteome Discoverer 2.

TMT reporter ions ratios were used for estimation of relative amount of each protein. Searches were done with the Human Uniprot reference database and a common contaminant database. Modification were set: peptide N terminus, lysine unimod and cystein unimod 39 as static, and methionine oxidation unimod and protein N-terminus acetylation unimod 1 as variable. The samples and related files are summarized in Table 2. The lysates in each series were adjusted to the same protein concentration with 1x SDS lysis buffer.

The results were presented as geometric means of fold change values with respect to control samples and the uncertainty was expressed as geometric standard errors. Beads were separated by brief centrifugation and washed two times with 1x Triton X lysis buffer. Finally, beads were resuspended in 1x Laemmli sample buffer 0. Samples were further processed according to the immunoblotting protocol described above.

Mass spectrometry data pre-processed with Proteome Discoverer 2. Differences in protein levels were estimated with moderated t-test statistics using the limma package Time-lapse movies documenting migratory phenotypes of the cells, uncropped immunoblot images, other supporting data referenced in the text, and complete results of differential transcript and protein level analyses were deposited in the Figshare 35 repository.

The different potential of the Src inhibitors to induce MAT could be attributed to their different effect on Src structure and localization To verify that the observed membrane blebbing is not due to initiation of apoptosis, we detected Caspase-3 in protein lysates from the cells in 3D collagen. While we easily detected the non-cleaved pro-form in all samples, only a faint signal of the active, cleaved form of Caspase-3 could be detected after a very long blot exposure in all samples with no significant differences Fig.

Numbers next to blots indicate average fold change, standard error both geometric and p-value of paired t-test. All results come from hour experiments. Quantitative analysis of cell migration. Track plots were generated using Chemotaxis Tool in ImageJ. Error bars — standard deviation. Representative results of three independent experiments. RNA sequencing of control and MAT-induced samples three pairs of independent biological replicates for each treatment yielded 31—46 million paired-end reads per sample.

Raw reads trimmed of adapter sequences were quality-checked with FastQC software The mapping metrics of STAR aligner showed an average paired read length of bases for inducible caRhoA samples and bases for dasatinib treatment series samples.

In average, the aligner uniquely mapped The complete mapping metrics is listed in Table 3. Transcriptomic and proteomic profiling of HT cells of mesenchymal and amoeboid migratory phenotype. We spent a great deal of time and real world testing with EGT sensors to make sure we were providing the best quality sensors possible without asking you to take out a mortgage to purchase these kits.

This search lead us to these high quality insulated K-Type thermocouples included with these kits, they are made from stainless steel, can withstand constant operating temperatures of over c for prolonged periods, and burst temps of up to c. These new EGT sensors cost 3x that of the old sensors supplied with these kits to produce, but we are dedicated here to providing the best quality sensors possible while still maintaining a reasonable price on the kits and feel the extra cost to achieve this level of quality on the EGT sensors was justified.

Another important factor which makes these new EGT sensors a cut above the rest is the majority of EGT sensors supplied with gauges are using single strand wires, these sensors are using multi strand. Now you may ask why is that better, well what we have discovered here after a year of playing around with various EGT sensors is this, heat is not the main killer of EGT sensors, vibration is. Having multiple strand wires in these sensors allows them to better cope with the harmonics and vibration of daily use, leading to less failures overall.

Then we have the high quality connectors, heat shielding, these sensors really are built to take a punishment. We were not going to mess around here as we know a lot of our customers use EGT, especially the 4x4 guys, just another way we have polished the v2 models of these kits. Now while this gauge system comes with all the sensors, there are two inputs that require you to source the signals from your vehicle as inputs into the control unit for the system to be able to display them on the 14in1 screen, these are RPM and Speed.

The first and quickest way to do this if you vehicle permits this is by using our OBDii cable which is sold separately and in most cases using this cable for the install will not only give you power for the units, but can pull both Speed and RPM from the OBD2 port directly to display on the gauge. RPM on the other hand more people will want, as there are functions in the digital multi gauge that can make use of an RPM input, such as some of the warning setups, you have shift lights on those as well.

We are pretty well versed here in how to obtain RPM for these systems, so when it comes time to install we can offer assistance in how to obtain an RPM signal. The v2 kits are now all built using high quality JST style connectors throughout, while most people never think quality of connectors means anything, using high quality JST style is just another way we have really polished the product. The cables slide in smoothly, the crimping and just overall build of the connectors on the control box, 10in1 gauge, sensor cables are top notch.

If you would like more information or have questions relating to this, please feel free to contact us. We have a range of accessories that can be bought along with this kit to make installation of the sensors or system simpler and neater.

Using these plates make the installation of the oil temp and oil pressure sensors much more straight forward, saves removing factory sensors, or having to tap and weld a fitting into your sump for oil temp as an example.

You will also need a sandwich plate adapter bolt for them which we also sell, before ordering, check which thread type your engine block uses for its oil filter to make sure one of the four adapters we sell will fit your motor. We also stock high quality stainless steel adapters, none of those cheap alloy units. Any questions in regards to these plates, feel free to contact us. Again this little product makes installing of the water temp sensor quick, simple and neat.

We currently stock three different sizes that cater to a large range of different cars, just like the sandwich plate adapters, before ordering check the size of your radiator hose to make sure which you need to order for your car, any questions give us a yell.

Brass cable gland for shielding against interference frequencies with a screening attenuation of up to dB. Contact with cable shield by means of spring washer. Brass cable gland for shielding against interference frequencies with a screening attenuation of up to 90 dB.

Multiple contact with cable shield by means of contact element. For increasing the class of protection for cable glands and blanking plugs recommended for use in Ex applications. Do you need extra threads, special laser markings or a different RAL colour? No problem! We put everything together according to your individual specifications, so that you end up with just the product that suits your needs. Completely pre-assembled, ready to use and naturally just as powerful, dynamic and forward-looking as our standard products.

Selecting products and devices for applications on a case-bycase basis can often be a time-consuming task. Kitting We can configure all individual connector components to meet your specific needs and provide you with a complete kit. Housing adaptation All of our housings can be designed to meet your specification needs. Pre-assembled connectors Looking for something extra? We are able to deliver ready-to-use connectors to meet your demands.

Laser marking of housings Individualised housing markings offer you endless opportunities. And it also means that you can take advantage of our comprehensive resources:. The start of a good partnership is always characterised by an intensive exchange of information to define the respective positions. Our contribution focuses on detailed advice with respect to:. And demanding standards guarantee a high level of quality every time. Our experience helps you create the foundation for good business and satisfied customers.

Production of terminal rails Terminal rails are manufactured from steel, stainless steel, aluminium or copper to suit the diverse applications. And we can produce terminal rails with elongated or round holes, or in other forms to suit your requirements exactly. Terminal rails are fitted with modular terminals or electronic products, prewired and marked according to your specification. Simply specify the colour and printing you require.

Special paint finishes and powder coating are also possible. Enclosures are adapted to suit the intended application exactly. You get a tailored, individual product and the quality is guaranteed by our adherence to demanding standards. Does your product require a special approval? Our accredited laboratory can test the complete product and confirm that its design complies with the standards!

With every delivery we document the corresponding approvals e. We bring the individual parts together according to your documents: whether for materials procurement, provision, or withdrawal from our stock.

All production processes and the entire range of qualified expertise are at our disposal: from hand assembly to SMD assembly. Our state-of-the-art production and testing facilities guarantee consistent quality. These are assembled and prewired according to your specification and are supplied ready to connect.

If required, we can also supply the finished enclosure with the heavy-duty connectors already integrated. An entire spectrum of application possibilities are available with our ConCept modular connector system. This modular system enables the flexible combination of diverse modules. Custom crimping and cabling is included on request! Do you prefer a personal touch? We can laser-label your company logo and article number onto our RockStars! In total compliance with your requirements.

However, we can also supply you with preprinted markers to match your specification. If required, we can also install the finished markers during assembly.

Equipment labelling Device markers are essential for marking your electrical installations. A wide range of different shapes, colours, materials and fixings — riveted, screwed or bonded with adhesive — are available from which to choose.

And a whole range of different fonts mean that we can handle every request. Integration of special accessories IIn some cases it is necessary to integrate special accessories.

And that includes the products of other manufacturers as well as our own. We shall also be happy to advise you on standard accessories such as hinges. And hence simplifies the ordering process enormously. Texts, borders, lines, graphics, barcodes, serial numbers and photographs are all possible. Besides product features and part numbers, it contains extensive additional information on all product groups. SG 10 BEZ. SF 10 BEZ. Canada W Interconnections Canada Inc.

Ecuador Elsystec S. Chile Felipe Bahamondes S. Narcis Monturiol Pol. Caspian Plaza, 5-th Floor 44 J. Jabbarly Str. Rua Garcia Lorca, V. Oktyabrskaya Str. Videnska ul. Indonesia PT. Ireland A. Haslam Ltd. Israel A. Shay Ltd. Iran TAF Co. Great Britain Weidmuller Ltd. Croatia Elektro Partner d. Asahi Seimei Bldg. Norway Siv. New Zealand Cuthbert S. O Box. United States W-Interconnections, Inc. Uruguay Rewo Uruguay S. Venezuela Somerinca C. III Makedonska brigada b.

Sciberras Str. Mexico W Interconnections, S. San Rafael Oriente Puebla, Pue. Service sesco-gex. Slovenia Elektrospoji d. Slovakia Elektris s. Thailand Pisanu Engineering Co. Mihrabad Cad. Taiwan Fittatek Co. Taiwan Eucan Enterprise Ltd. Linh Kim Hai Co,. Index We cannot guarantee that there are no mistakes in the publications or software provided by us to the customer for the purpose of making orders.

We try our best to quickly correct errors in our printed media. For this purpose we list corrections, which have been identified after publishing printed media, on our web site at www. This Internet-database is part of our catalogue. All orders are based on our general terms of delivery, which can be reviewed on the websites of our group companies where you place your order.

On demand we can also send the general terms of delivery to you. When you are able to look at the gauge again, hold the peak recall button and the gauge will show you the first 15 seconds of data from that mode, as well as RPM scaled onto the bar graph readout.

For pre vehicles, you must enable code reading in the configuration menu. Code reading at startup is disabled by default on pre model year cars because the code reading process on older vehicles can take up to 10 seconds, and it would delay startup. C option. Use this to clear codes. Display Options: d. Enter config with car OFF to re-enable from valet. Units setting: En. Boost Pressure Resolution: Pr0. Vacuum Bar graph: U. Bar graph indicates vacuum by moving right to left. Bar graph does not indicate vacuum.

Shift Light: SL. Boost Reading Selection: bSt. Analog Input Configuration see the analog inputs section for details : A1. Car Selection: CAr.

Allows gauge to read only Boost and battery voltage without communicating on the vehicle network. Requires add-on analog boost sensor. Code Clearing: Clr. HOLD the left button until screen goes blank or "Clrd" is displayed to clear codes. See Code Reading and Clearing section above for details. You can also double-check this by unplugging the hose from your analog sensor while the car is running. If the gauge shows "0. Advanced Protocol Setting: Sh.

Vehicle specific software protocol adjustments See configuration list for details, use Sh. A unless otherwise directed. The multi-gauge harness has 3 linear analog inputs, 2 of which can be used as universal displays for ANY v sensor with a linear ascending output. The 3rd is used for the Analog Boost Sensor. Do not hook up a device that sends more than 5 volts or you WILL damage the control box.

To enable an analog input, enter the config menu and set the A1 input to Y for yes. After doing this, 3 new options will appear:. DP , analog 1 decimal point position. The gauge will show on the screen, and tapping the first button will move the decimal point. L is the bargraph start, H is the bargraph end.