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. 2021 Mar 30;10(4):755. doi: 10.3390/cells10040755

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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SHBG protects against PA-induced endoplasmic reticulum (ER) stress. HepG2 cells were incubated with SHBG in two concentrations (50 or 100 nM) before exposition to PA for 24 h. ER localization in cells was visualized with ER-TrackerTM Green (A). mRNA levels of IRE1A (B), ATF6 (C), CHOP (D), and HSPA5 (E) were determined using the RT-qPCR method. The phosphorylation of eIF2α at Ser51 and IRE1α at Ser724, as well as the cleavage activation of ATF6 were estimated using the Western blot technique (F). Relative intensities of ATF6 full length (G), ATF6 cleaved (H), phosphorylated IRE1α (I), and eIF2α (J) were calculated with Image Lab software after normalization to β-actin as a control housekeeping protein. Intracellular localization of XBP1 investigated with confocal microscopy (K). Expression levels (L) of spliced and unsliced XBP1 established by RT-qPCR followed by gel electrophoresis. The results are presented as mean ± SEM; n = 6, * p < 0.05, ** p < 0.01, *** p < 0.001.

SHBG protects against PA-induced endoplasmic reticulum (ER) stress. HepG2 cells were incubated with SHBG in two concentrations (50 or 100 nM) before exposition to PA for 24 h. ER localization in cells was visualized with ER-TrackerTM Green (A). mRNA levels of IRE1A (B), ATF6 (C), CHOP (D), and HSPA5 (E) were determined using the RT-qPCR method. The phosphorylation of eIF2α at Ser51 and IRE1α at Ser724, as well as the cleavage activation of ATF6 were estimated using the Western blot technique (F). Relative intensities of ATF6 full length (G), ATF6 cleaved (H), phosphorylated IRE1α (I), and eIF2α (J) were calculated with Image Lab software after normalization to β-actin as a control housekeeping protein. Intracellular localization of XBP1 investigated with confocal microscopy (K). Expression levels (L) of spliced and unsliced XBP1 established by RT-qPCR followed by gel electrophoresis. The results are presented as mean ± SEM; n = 6, * p < 0.05, ** p < 0.01, *** p < 0.001.