Figure 5
From: Tumor-derived neomorphic mutations in ASXL1 impairs the BAP1-ASXL1-FOXK1/K2 transcription network
Tumor-derived ASXL1 mutant regulates glucose metabolism, oxygen sensing, and JAK-STAT3 signaling pathways. (A) The mRNA expression of indicated genes in FOXK1 and/or FOXK2 knockout K562 cell pools, as determined by qRT-PCR. (B) The mRNA expression of indicated genes in ASXL1+/N590 and ASXL1+/− K562 clones, as determined by qRT-PCR. (C) TXNIP protein level in ASXL1+/N590 and ASXL1+/− clones, as determined by Western blot. (D) Glucose uptake in ASXL1+/N590 and ASXL1+/− clones, as measured using a Glucose Assay Kit as described in the “MATERIALS AND METHODS” section. (E) Relative concentration of lactate, ATP, and ADP in ASXL1+/N590 and ASXL1+/− K562 clones, as measured by LC–MS/MS as described in the “MATERIALS AND METHODS” section. (F) The protein levels of HIF-1α in ASXL1+/N590 and ASXL1+/− K562 clones under the culture conditions of normoxia or hypoxia (1% and 8% O2, exposure for 48 h). (G) The mRNA expression of HIF-1α target genes in ASXL1+/N590 and ASXL1+/− K562 clones under the culture conditions of normoxia or hypoxia (1% O2, exposure for 48 h). (H) JAK2, STAT3, and their site-specific phosphorylation levels in ASXL1+/N590 and ASXL1+/− clones. (I) The mRNA expression of STAT3 target genes in ASXL1+/N590 and ASXL1+/− clones, as determined by qRT-PCR. Asterisks denote statistical significance with two-tailed Student’s t-test (B, D, E, G, I) or one-way ANOVA (A). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 for the indicated comparison; ns = not significant