Boxes show three individual areas that have been and expanded, with indicated channels separated. Arrowheads mark p21 + /CD80 + and IRF1 + /PD-L1+ cells ; γH2AX + /CD80+ cells and γH2AX/PD-L1 negative cells ; p-Stat3+/PD-L1+ and p-Stat3 + /CD80+ cells .Untreated tumor stained and imaged in parallel. Scale bar=40 µm. qPCR and IF were repeated at least twice and representative data are shown.
CRISPR-Cas9 was used to generate a p53 knockout clone and a non-edited control of 4226 cell line.p53 knockout clone 2 of the 4226 cell line was untreated or treated with doxorubicin as indicated. Over 1 to 5 days, these cultures were then exposed to rIFNγ for 24 h ) or not , followed by immunoblot for CD80, PD-L1, Stat1, and IRF1 on a total of 2 membranes, which were then blotted for actin.
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