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. 2023 Feb 1;133(3):e160456. doi: 10.1172/JCI160456

Figure 1. MIB2 regulates membrane PD-L1 levels in tumor cells.

Figure 1

(A) Volcano plot showing the E3 ligases identified from FACS. MIB2 is indicated in red, and STUB1 and BTRC are indicated in blue. (B) Immunoblotting (IB) analysis of PD-L1 levels in MIB2-KO B16-F10 and MC38 cells. (C and D) FACS analysis of membrane PD-L1 levels in (C) B16-F10 and (D) MC38 cells. (E and F) IB analysis of PD-L1 protein levels in whole-cell extract (WCE) and membrane fractions (Mem) from (E) B16-F10 and (F) MC38 cells. (G and H) Immunofluorescence analysis of PD-L1 in B16-F10 and MC38 cells. (G) Representative images of green fluorescence–labeled PD-L1. Scale bar: 10 μm. (H) Quantitative analysis of membrane- and cytoplasmic-expressed PD-L1 (n = 5). Cyto, cytoplasm. ***P < 0.001 by 1-way ANOVA test with Dunnett’s multiple comparisons test (C, D, and H).