Background
Methods
Experimental groups, workflow, and study design of TFRE profiling
HUVEC cultivation
Nuclear protein extraction
Enrichment of TFs in nuclear protein extracts using catTFRE
Nano-LC–MS/MS analysis for protein identification and label-free quantification
Selection of candidates for key TFs in HUVEC apoptosis
Western blot (WB) analysis
Real time quantitative PCR (RT-qPCR)
Molecules | Forward | Reverse |
---|---|---|
MAFG | TCCAGGGTACTGACCTGCTC | GTTTCCTTTATTGGGGGTCG |
MAFF | TGCCCAGGTCCCATTTCTC | GGCCCACGAAGGGAATGT |
β-actin | CCTAGAAGCATTTGCGGTGG | GAGCTACGAGCTGCCTGACG |
MAFF or MAFG overexpression (OE) plasmid construction and transfection of SV40T-transformed HUVECs
RT-qPCR and WB validation of MAFF or MAFG OE
Molecules | Forward | Reverse |
---|---|---|
MAFG | AAGGCCTTGAAGGTGAAGCG | CCTTCTGCTTCTCCAGCTCC |
MAFF | GAGGAGCTGCAGAAGCAGAA | TGACGATGGTGATGACGCTG |
GAPDH | ACAGCAACAGGGTGGTGGAC | TTTGAGGGTGCAGCGAACTT |