Fig. 6: PD-L1 K263 acetylation abrogates its unclear translocation.
From: Nuclear PD-L1 promotes EGR1-mediated angiogenesis and accelerates tumorigenesis
a Western blot analysis of anti-PD-L1 IPs derived from MUM2B, MEL290 and A375 cells to detect PD-L1 acetylation levels. b Western blot images showing the protein levels of EP300 and HDAC2 in MUM2B, MEL290 and A375 cells. c Western blot analysis of anti-PD-L1 IPs derived from MUM2B and MEL290 cells treated with the indicated constructs to detect PD-L1 acetylation levels. d, e Western blot analysis of cytoplasmic/membrane (C/M) and nuclear (N) fractions of PD-L1 in MUM2B (d) and MEL290 (e) cells transduced with si-EP300. f ChIP-qPCR assay of the PD-L1 status at the EGR1 promoter in MUM2B and MEL290 cells transduced with si-EP300. g Western blot images showing the protein levels of EGR1 and VEGFA in MUM2B and MEL290 cells transduced with si-EP300. h, i Western blot analysis of cytoplasmic/membrane (C/M) and nuclear (N) fractions of PD-L1 in MUM2B (h) and MEL290 (i) cells treated with 50 μM HDAC2i (santacruzamate A) for 24 h. j ChIP-qPCR assay of the PD-L1 status at the EGR1 promoter in MUM2B and MEL290 cells treated with 50 μM HDAC2i for 24 h. k Western blot images showing the protein levels of EGR1 and VEGFA in MUM2B and MEL290 cells treated with 50 μM HDAC2i for 24 h. l Western blot analysis of cytoplasmic/membrane (C/M) and nuclear (N) fractions of PD-L1 in MUM2B cells treated with the indicated constructs. m Statistical analysis of nPD-L1 levels in MUM2B cells treated with the indicated constructs. n = 3. Data are presented as means ± SD. Two-tailed unpaired Student’s t-test. n IF staining of PD-L1 in MUM2B cells treated with the indicated constructs. Left panel, representative IF staining images. Scale bars, 10 µm. Right panel, co-localization analysis of PD-L1 and DAPI by ImageJ software. o Statistical analysis of nPD-L1 fluorescence intensity (%) in Fig. 6n. n = 20. Data are presented as means ± SD. Two-tailed unpaired Student’s t-test. p Western blot images showing the protein levels of EGR1 and VEGFA in MUM2B cells treated with the indicated constructs. q Schematic diagram showing that PD-L1 nuclear translocation was mediated by HDAC2-dependent PD-L1 K263 deacetylation.
