Figure 1

Hypoxia reduces p62 mRNA and protein levels. (A) Hep3B cells were incubated under hypoxia (O₂ = 1%) for 2, 4, 6, 8, and 12 h. The protein concentrations of p62, HIF-1α, and β-actin were examined by immunoblotting. (B) Hep3B cells were incubated under hypoxia (O₂ = 1%) for the indicated period. The mRNA levels of p62 were assessed by RT-PCR and quantified relative to β-actin. (C and D) Hep3B cells were incubated under hypoxia (O₂ = 1%) or treated with 100 µM dipyridyl or 100 µM CoCl₂ under normoxia for 12 h. The protein concentration (C) and mRNA abundance (D) of p62 were measured by immunoblotting and RT-PCR, respectively. (E and F) Hep3B cells were transfected with siRNA-Control, siRNA-HIF-1α (HIF-1α K.D.), or siRNA-HIF-2α (HIF-2α K.D.) and were incubated under hypoxia (O₂ = 1%, 12 h). The protein concentration (E) and mRNA abundance (F) of p62, HIF-1α, HIF-2α, and β-actin were measured by immunoblotting and RT-PCR, respectively. Data are the mean ± S.D. of three replicates. N.S. not significant, *p < 0.05, **p < 0.01, ***p < 0.001 versus the indicated group, calculated with a one-way ANOVA with Dunnett’s post-hoc test (for figures A–D), a one-way ANOVA with Tukey’s post-hoc test (for figure E), or a multivariate one-way ANOVA with Tukey’s post-hoc test (for figure F).