Fig. 1
Increased HIF-1α expression in activated B cells. a Quantitative RT-PCR analyses of Hif1a and Hif2a in wild-type (WT) splenic B cells stimulated with lipopolysaccharide (LPS) or anti-IgM at indicated time points (n = 4 at each time point). Values at 0 h were set as 1. b Expression of HIF-1α and HIF-2α in WT splenic B cells stimulated with LPS or anti-IgM at indicated time points (n = 3 experiments in duplicate). c Western blot and densitometry analysis of phospho-STAT3 Ser727 (pSTAT3727), phospho-STAT3 Tyr705 (pSTAT3705), total STAT3, phospho-ERK (pERK), and total ERK in WT splenic B cells after stimulation with anti-IgM at indicated time points (n = 3 experiments in duplicate). d Western blot and densitometry analysis of HIF-1α, pSTAT3727, and pERK in anti-IgM-stimulated B cells with or without STAT3, ERK, or AKT inhibitors treatments for 4 h (n = 3 experiments in duplicate). e Scheme of Hif1a promoter indicating the potential STAT3 binding site position and enrichment of pSTAT3727 on Hif1a promoter in splenic B cells 4 h after stimulation with anti-IgM or medium (Med) (n = 4 for all groups). Data are shown as mean ± s.e.m. Pictures are representative of three (a–d) or four (e) independent experiments. *P < 0.05; **P < 0.01, and ***P < 0.001 (two-tailed unpaired Student’s t-test) (see also Supplementary Figure 1)
