Figure 9. BHLHE40 promotes LPS-induced inflammatory and glycolytic gene expression through HIF1α.
(A) RAW264.7 cells were transfected with pcDNA3 or pcDNA3-ΔHif1α plasmid. Total protein extracts from these cells were evaluated for HIF1α expression by western blots.
(B and C) RAW264.7 cells were transfected with Bhlhe40-specific siRNA or/and ΔHif1α plasmid. These cells were stimulated with 100 ng/ml LPS for 4 hours. Total RNA from these experiments were evaluated for expression of Ptgs2, Nos2, Il12a, Cxcl3 (B) and Glut1, Pfkfb3, Pgk1, Ldha (C) by RT-qPCR (n=4). Actin and 36B4 were used as housekeeping genes for western blot and RT-qPCR analyses, respectively. (D) BHLHE40 promotes macrophage inflammatory gene programs, hypoxia response, glycolytic gene expression, and pathogenic inflammatory responses by elevating HIF1α expression. All values are reported as mean ± SD. Data were analyzed by ANOVA followed by Bonferroni post-testing. N.S., not significant; ** p < 0.01; ***F p < 0.001.