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. 2020 Nov 28;253(2):209–224. doi: 10.1002/path.5574

Figure 4.

Figure 4

FERM2 regulates EC gene expression by preventing their mRNA decay via interacting with DNA/RNA binding protein YBX1. (A) VWF and CDH5 gene promoter activity was not regulated by FERM2. (B) VWF and CDH5 mRNA degradation was increased by FERMT2 inhibition. HUVECs infected with a non‐target (sh‐NT) or FERMT2 gene‐specific shRNA (sh‐FERM2) lentivirus were treated with the transcription inhibitor (actinomycin D, ActD, 1 μg/ml) for the indicated times. The VWF/CDH5 mRNA arbitrary unit at 0 h for both groups was set as 1.0, and at other time points was calculated accordingly. (C) Proximity ligation assays (PLAs) showed the in situ protein interactions of FERM2 with YBOX1 in HUVECs. (D) RNA immunoprecipitation (RIP) assays were conducted in HUVECs infected with sh‐NT or sh‐FERM2 lentivirus using antibody against YBOX1 or normal IgG, respectively. Immunoprecipitated RNAs were subjected to RT‐qPCR with the indicated gene primers. The data presented here are representative images (C) or mean ± SEM (A, B, D) of five (n = 5) independent experiments. *p < 0.05, **p < 0.01 (versus sh‐NT); # p < 0.05, ## p < 0.01 (versus 0 h or Intron). Two‐way ANOVA with a Tukey's post hoc test (B) or Student's t‐test (D) was used for statistical analysis, respectively.