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. 2022 May 21;163(7):bqac070. doi: 10.1210/endocr/bqac070

Figure 2.

Figure 2.

Hypoxia inhibits expression of transcription factors NRF2 and C/EBPβ in cultured hTSCs. hTSCs cultured in SynT differentiation medium containing forskolin in 20% O2 or 2% O2 for 1, 3, or 5 days were harvested for analysis by RT-qPCR and western blotting. mRNA (A) and nuclear protein levels (D, E) of NRF2 increased markedly during culture of hTSCs in 20% O2 and were inhibited in response to culture in a hypoxic (2% O2) environment after 3 and 5 days of culture. (A, D, E). Keap1 protein expression was reduced during SynT differentiation after 5 days of culture in 20% oxygen and remained increased at levels similar to those of undifferentiated CytT after 5 days of culture in 2% O2 (D, F). Expression of C/EBPβ at both mRNA (B) and protein (G, H) levels was markedly increased after 3 and 5 days of culture in SynT differentiation medium in 20% O2 and was inhibited in response to culture in 2% O2. Shown are representative images of immunoblots (D, G) and quantifications of the scans of blots (E-I). Values are the means ±SEM of data from 3 to 5 independent experiments; *P < 0.05; **P < 0.01; ***P < 0.001 vs 0 d; #P < 0.05; ##P < 0.01; ###P < 0.001 vs 20% O2 at 5 days.