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. Author manuscript; available in PMC: 2022 Oct 1.
Published in final edited form as: FASEB J. 2021 Oct;35(10):e21940. doi: 10.1096/fj.202100944R

Figure 7. BHLHE40 boosts HIF1α and pro-inflammatory gene expression in macrophages.

Figure 7.

(A and B) The Lyz2cre/cre and Bhlhe40fl/fl:Lyz2cre/cre mice were challenged with intratracheal delivery of saline or zymosan (n=3). Macrophages derived from bronchoalveolar lavages were evaluated for HIF1α protein levels by western blot (A). The total HIF1α levels were quantified by densitometry analysis (B). (C-F) Lyz2cre/cre and Bhlhe40fl/fl:Lyz2cre/cre mice PMs were stimulated with 100 ng/ml LPS (C) or exposed to hypoxic conditions (E) for 4 hours. Total protein extracts were evaluated for the expression of HIF1α by western blot (n=3). The total HIF1α levels were quantified by densitometry analysis (D and F). (G) RAW264.7 cells were transfected with Hif1α-specific siRNA or/and pcDNA3-BHLHE40 plasmid. These cells were stimulated with 100 ng/ml LPS for 4 hours. Total RNA from these experiments were evaluated for expression of Ptgs2, Fpr1, Mmp13, and Cd14 by RT-qPCR (n=4). Actin and 36B4 were used as housekeeping genes for western blot and RT-qPCR analyses, respectively. Values are reported as mean ± SD. Data were analyzed by ANOVA followed by Bonferroni post-testing. *p < 0.05, ** p< 0.01 and ***p < 0.001.