Fig. 6: IR induces PD-L1 foci, co-localization and direct interaction with Ku80 in the nucleus. | Nature Communications

Fig. 6: IR induces PD-L1 foci, co-localization and direct interaction with Ku80 in the nucleus.

From: PD-L1 deglycosylation promotes its nuclear translocation and accelerates DNA double-strand-break repair in cancer

Fig. 6

a Co-localizations of PD-L1 with Ku80 or γH2AX were analyzed in H460 cells following IR exposure by immunofluorescence with pre-extraction using CSK plus RNase A (CSK + R). b H460 cells were treated with IR, followed by isolation of cytoplasmic and nuclear fractions. Co-IP experiments using PD-L1 antibody were performed in cytoplasmic and nuclear fractions, followed by western blot. IgG and lysate were used as controls. **PD-L1: glycosylated form, *PD-L1: nonglycosylated form. α-tubulin as cytoplasmic marker, and PCNA as nuclear marker, for purity of each fraction. c PD-L1/Ku80 interactions were analyzed by PLA following IR (4 Gy) exposure in H460 cells. PLA signals were detected by fluorescence microscopy as discrete spots. IgG, single PD-L1 antibody alone or single Ku80 antibody alone was used as negative control. After PLA, immunofluorescence staining using α-tubulin antibody was performed as phase contrast imaging. Error bars represent ± s.d., n = 10 per group. ***P < 0.001, by two-tailed t test. Source data are provided as a Source Data file.

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