What is the best way to identify a cell line that expresses VEGFR3 exclusively, or with very low expression of VEGFR2 and VEGFR1?

Nouhaila Elmoumen

To find a cell line expressing VEGFR3 and low VEGFR1 and VEGFR2:

1. **Research:** Look up existing studies or databases for suitable cell lines.

2. **Database Check:** Use CCLE or GDSC for gene expression data.

3. **Analyze Data:** Select cell lines with low VEGFR1 and VEGFR2, high VEGFR3.

4. **Validation:** Confirm gene expression using qPCR, western blotting, or staining.

5. **Test Peptide:** Assess peptide binding and downstream effects.

6. **Control:** Include control treatments for comparison.

7. **Replicate:** Repeat experiments for reliable results.

Saif Wahid

Identifying a cell line that expresses VEGFR3 exclusively or with very low expression of VEGFR2 and VEGFR1 can be challenging, as many cell lines express multiple VEGF receptors. However, here are some approaches to help you identify suitable cell lines:

Cell surface protein expression analysis: Use flow cytometry or fluorescence microscopy to analyze the cell surface expression of VEGFR1, VEGFR2, and VEGFR3 in various cell lines. This will help you identify cell lines with high expression of VEGFR3 and low expression of VEGFR1 and VEGFR2.

qRT-PCR analysis: Perform quantitative reverse transcription polymerase chain reaction (qRT-PCR) to evaluate the mRNA expression levels of VEGFR1, VEGFR2, and VEGFR3 in different cell lines. This will give you an idea about the relative expression levels of these receptors in each cell line.

Database mining: Utilize online databases such as ArrayExpress, Gene Expression Omnibus (GEO), or Cancer Cell Line Encyclopedia (CCLE) to explore gene expression profiles of various cell lines. You can use tools like GEO2R or limma to compare the expression levels of VEGFR1, VEGFR2, and VEGFR3 across different cell lines.

Knockdown or knockout experiments: Consider performing knockdown or knockout experiments using CRISPR-Cas9 or shRNAs to specifically silence VEGFR1 and VEGFR2 in cell lines that express these receptors. This will allow you to evaluate the effect of VEGFR3 expression in the absence of VEGFR1 and VEGFR2.

Use cell lines with known VEGF-A/VEGFR interactions: Some cancer cell lines, such as HUVEC (human umbilical vein endothelial cells) and HMVEC (human microvascular endothelial cells), are known to express high levels of VEGFR3 and have been shown to interact with VEGF-A. You may also consider using cell lines that have been engineered to overexpress VEGFR3.

Screening assays: Develop or utilize existing screening assays that measure the binding affinity of your peptide to VEGFR1, VEGFR2, and VEGFR3. This will enable you to identify cell lines with high binding affinity for your peptide, indicating strong expression of VEGFR3 and potentially low expression of VEGFR1 and VEGFR2.

Combinatorial approaches: Combine the above approaches to increase the confidence in identifying cell lines that meet your criteria. For example, you could first narrow down your options by analyzing gene expression profiles from online databases, followed by validating the expression levels using qRT-PCR or flow cytometry.

Validation with orthogonal methods: Once you have identified potential cell lines, validate their expression profiles using orthogonal methods such as western blotting or immunofluorescence staining. This will provide further confirmation of VEGFR expression levels and help rule out any discrepancies due to technical variations.

Test your peptide: Finally, test the binding specificity of your peptide on the shortlisted cell lines using techniques such as flow cytometry, immunoprecipitation, or surface plasmon resonance. This will help you determine the most suitable cell line(s) for your studies.

Remember that it is crucial to carefully evaluate and validate the expression profiles of VEGFR1, VEGFR2, and VEGFR3 in the selected cell lines to ensure the accuracy and reliability of your results.

Hello fellow researcher .. What is the meaning of your name?

Can we classify HV and LV lines based on line length?

Does the pressure(-600 mmHg) affects pH?

After a reference from Int J Body Compos Res, can any body help?

Do anyone here can help us in the field of molecular docking?

Analysis of CRD two factor using R studio ?

Which is better medicine or food in obesity?

How could I take into consideration the weights of observation in a system of seemingly unrelated regressions (SUR) in rstudio?

What is the exact mechanism of the antimicrobial effect of local anesthetics?

Can anyone share his experience regards Smoothing in origin?

Can we classify HV and LV lines based on line length?

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

No title

Issues with Permeabilization during Flow Cytometry?

Which cell line should we use to perform biological dosimetry experiments?

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

What could be some reasons why unstimulated monocytes are getting activated?

What is the ideal efficiency for a qPCR?