Fig. 5: Identification of interaction between SET8 and KLF4. | Cell Death & Disease

Fig. 5: Identification of interaction between SET8 and KLF4.

From: Monomethyltransferase SET8 facilitates hepatocellular carcinoma growth by enhancing aerobic glycolysis

Fig. 5

a 97H cells were infected with adenovirus-expressing GFP (Ad-GFP) or SET8 tagged with Flag epitope (Ad-SET8). Cells were harvested, and nuclear extracts were sequentially immunoprecipitated with Anti-FLAG (M2 agarose). The SET8-associated proteins were detected by SDS/PAGE and silver staining. b HEK293T cells were ectopically expressed with the indicated plasmids (Flag-SET8 and HA-KLF4). Extracts were immunoprecipitated with the antibody against Flag tag (IP: Flag-SET8/IB: KLF4) or KLF4 (IP: HA-KLF4/IB: Flag-SET8). c Validation of SET8–KLF4 interaction in vivo. Endogenous interaction between SET8 and KLF4 was observed in HCC cells measured by immunoprecipitation. d Colocalization of SET8 and KLF4 in HCC cells detected by confocal microscopy. e mRNA level of KLF4 detected in SET8-overexpressing cells. f, g Analysis of extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) in M3 (left) and 97H (right) cells with either KLF4 overexpression or control vector. h, i The expression levels of genes involved in glucose metabolism in the indicated HCC cell lines with either KLF4 overexpression or control vector detected by real-time PCR and western blot analysis. Data are shown as mean ± SD of three independent experiments. *P < 0.05

Back to article page