Fig. 4: Nuclear PD-L1 facilitates angiogenesis through EGR1.
From: Nuclear PD-L1 promotes EGR1-mediated angiogenesis and accelerates tumorigenesis
a HUVEC migration assay that investigates the effect of EGR1 re-expression in PD-L1-silenced cells on the migration ability of HUVECs. Scale bars, 100 µm. b Statistical analysis of the HUVEC migration assay. n = 3. Data are presented as means ± SD. Two-tailed unpaired Student’s t-test. c HUVEC tube formation assay that investigates the effect of EGR1 re-expression in PD-L1-silenced cells on HUVEC tubule formation. Scale bars, 25 µm. d Statistical analysis of the HUVEC tube formation assay. n = 3. Data are presented as means ± SD. Two-tailed unpaired Student’s t-test. e CAM assay that investigates the effect of EGR1 re-expression in PD-L1-silenced cells on the blood vessel density of CAM. f Statistical analysis of the CAM assay. n = 5. Data are presented as means ± SD. Two-tailed unpaired Student’s t-test. g Representative bioluminescent images of UM orthotopic xenografts generated by PD-L1-silenced cells that re-express EGR1. h Statistical analysis of the bioluminescent signals in UM orthotopic xenografts. n = 5. Data are presented as means ± SD. Two-tailed unpaired Student’s t-test. i Statistical analysis of the tumor weight. n = 5. Data are presented as means ± SD. Two-tailed unpaired Student’s t-test. j Representative images of H&E staining of UM orthotopic xenografts generated by PD-L1-silenced cells that re-express EGR1. k IF images of CD31 in UM orthotopic xenografts constructed by EGR1 re-expression in PD-L1-silenced cells. Scale bars, 40 µm. l Statistical analysis of CD31 expression in UM orthotopic xenografts. n = 5. Data are presented as means ± SD. Two-tailed unpaired Student’s t-test. m–o Gene expression correlation analysis of PD-L1 and EGR1 (m), PD-L1 and VEGFA (n), EGR1 and VEGFA (o) in UM patients using TCGA database. p, q Kaplan–Meier analysis of the overall survival of UM patients with low and high EGR1 (p) and VEGFA (q) expression levels using TCGA database. r Schematic diagram showing that nPD-L1 facilitates angiogenesis in UM through transcriptional activation of EGR1.
