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. 2022 Jan 20;9(8):2104055. doi: 10.1002/advs.202104055

Figure 2.

Figure 2

PKM2 interacts with FACT and enhances the interaction between FACT and nonchromatin bound γH2AX. Immunoprecipitation (IP) and immunoblot (IB) analyses were performed with indicated antibodies. Data are representative of at least three independent experiments. A,B) U251 cells were transfected with A) Flag‐SPT16 or B) SFB‐SSRP1 and treated with etoposide (200 × 10–6 m) for indicated time. Whole‐cell lysates were prepared without sonication. C) U251 cells with or without SSRP1 depletion were treated with or without etoposide (200 × 10–6 m, 1 h). The chromatin fraction was prepared. D,E) U251 cells stably expressing shNT or shPKM2 were infected with the lentivirus expressing D) SFB‐SSRP1 or E) SFB‐SPT16 and treated with or without etoposide (200 × 10–6 m, 1 h). The nonchromatin‐bound fraction was prepared. F,G) PKM2‐depleted U251 cells were rescued with rPKM2 WT or K305Q and infected with the lentivirus expressing F) SFB‐SSRP1 or G) SFB‐SPT16. The cells were treated with or without etoposide (200 × 10–6 m, 1 h) and the nonchromatin‐bound fraction was prepared. H) PKM2‐depleted U251 cells were rescued with rPKM2 WT or K305Q followed by etoposide treatment (200 × 10–6 m, 1 h). Chromatin fraction (left panel) and nonchromatin‐bound fraction (right panel) were prepared.