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

Figure 7. Ubiquitination by MIB2 is required for the PD-L1 and RAB8 interaction and exocytosis.

Figure 7

(A) Coimmunoprecipitation (Co-IP) and immunoblotting (IB) analysis of endogenous PD-L1 and RAB8 in B16-F10 cells. (B and C) Proximity ligation assay (PLA) analysis of the PD-L1 and RAB8 interaction in MC38 cells. (B) Representative images. Scale bar: 10 μm. (C) Quantitation. Lines within the boxes denote median values; the tops of boxes represent the upper quartile (75th percentile), and bottoms of boxes represent the lower quartile (25th percentile); and widths denote cell densities. (D) Co-IP and IB analysis of endogenous PD-L1 and exocyst components in B16-F10 cells. (E) Co-IP analysis of the PD-L1 (WT or K136R) and RAB8 interaction in PD-L1–KO B16-F10 cells transfected with the indicated constructs. (F) IB analysis of PD-L1 in the whole-cell extract (WCE) and isolated Golgi from RAB8-silenced MC38 cells. ***P < 0.001, by 1-way ANOVA among 3 groups (B and C). Data are shown as the mean ± SEM.