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. 2021 Apr 13;9:665089. doi: 10.3389/fcell.2021.665089

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Copyright © 2021 Yu, Wu, Li, Zhou, Shen, Gao, Lin, Dai, Ou, Liu, Huang, Zheng and Sun.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Both cytoplasmic and nuclear BMI1 are required for steroidogenesis. (A) The assessment of testosterone levels in MLTC-1 cells treated as indicated for 48 h. PTC-209, N-acetylcysteine (NAC), pcDNA3.0-Bmi1-flag, and pcDNA3.0-Bmi1-△NLS2-flag were used at the concentrations of 5 μM, 500 μM, 1 μg/mL, and 1 μg/mL, respectively. Testosterone levels were determined by ELISA after treatment with 1 IU/mL human chorionic gonadotropin (hCG) for 6 h. (B) Immunofluorescence staining for FLAG in MLTC-1 cells transfected with the pcDNA3.0-flag empty vector (EV), pcDNA3.0-Bmi1-flag, and pcDNA3.0-Bmi1-△NLS2-flag for 48 h. All constructs were used at the concentration of 1 μg/mL. Scale bar, 20 μm. *p < 0.05, ***p < 0.001.