Journal: Scientific reports

Article Title: ERS regulates endometrial epithelial cell autophagy through XBP1s-mediated activation of the PI3K/AKT pathway.

doi: 10.1038/s41598-024-84461-6

Figure Lengend Snippet: Fig. 5. XBP1s knockdown reversed the effect of TG on gEECs autophagy. The experimental treatment conditions were as follows: pretreatment with TG for 2 h, followed by rapamycin treatment 24 h. (A-D) The relative protein expression of SQSTM1, ATG5, and the LC3II/LC3I were analyzed using western blotting and were quantified by densitometry. (E-F) The relative SQSTM1 and ATG5 levels were quantified through RT- qPCR. Data is represented as the means ± SEM of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 vs. control group. #, P < 0.05; ##, P < 0.01; ###, P < 0.001 vs. other group.

Article Snippet: The medium was replaced with fresh medium and the following treatments were initiated: (1) rapamycin (50 nM; TargetMol, Boston, MA, USA) for 3 h, 6 h, 12 h, and 24 h; (2) pretreatment with TG (Thapsigargin; 50 nM; an ERS activator; TargetMol) or 4-PBA (4-Phenylbutyric acid; 1 mM, an ERS inhibitor; Abcam, Cambridge, UK) or SC79 (10 μM; a PI3K/AKT pathway activator; TargetMol) or LY294002 (10 μM, a PI3K/AKT pathway inhibitor; TargetMol) for 2 h, followed by rapamycin treatment.

Techniques: Knockdown, Expressing, Western Blot, Quantitative RT-PCR, Control

Journal: Scientific reports

Article Title: ERS regulates endometrial epithelial cell autophagy through XBP1s-mediated activation of the PI3K/AKT pathway.

doi: 10.1038/s41598-024-84461-6

Figure Lengend Snippet: Fig. 8. Effect of PI3K/AKT pathway inhibition by LY294002 on overexpression of XBP1s. The experimental treatment conditions were as follows: pretreatment with LY294002 for 2 h, followed by rapamycin treatment 24 h. (A-D) Quantification of SQSTM1, ATG5, and the LC3II/LC3I band intensities from three independent experiments as determined using densitometric analysis. (E-F)SQSTM1 and ATG5 levels were detected using RT-qPCR. (G) Immunofluorescence images of LC3B expression in gEECs. Representative images of three independent experiments are shown. Scale bar = 10 μm. Data is represented as the means ± SEM of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 vs. control group. #, P < 0.05; ##, P < 0.01; ###, P < 0.001 vs. other group.

Article Snippet: The medium was replaced with fresh medium and the following treatments were initiated: (1) rapamycin (50 nM; TargetMol, Boston, MA, USA) for 3 h, 6 h, 12 h, and 24 h; (2) pretreatment with TG (Thapsigargin; 50 nM; an ERS activator; TargetMol) or 4-PBA (4-Phenylbutyric acid; 1 mM, an ERS inhibitor; Abcam, Cambridge, UK) or SC79 (10 μM; a PI3K/AKT pathway activator; TargetMol) or LY294002 (10 μM, a PI3K/AKT pathway inhibitor; TargetMol) for 2 h, followed by rapamycin treatment.

Techniques: Inhibition, Over Expression, Quantitative RT-PCR, Immunofluorescence, Expressing, Control

Journal: Scientific reports

Article Title: ERS regulates endometrial epithelial cell autophagy through XBP1s-mediated activation of the PI3K/AKT pathway.

doi: 10.1038/s41598-024-84461-6

Figure Lengend Snippet: Fig. 9. Effect of PI3K/AKT pathway activation by SC79 on knockdown of XBP1s. The experimental treatment conditions were as follows: pretreatment with SC79 for 2 h, followed by rapamycin treatment 24 h. (A-D) Relative protein expression levels of SQSTM1, ATG5, and the LC3II/LC3I were analyzed and determined using western blotting. (E-F) The relative mRNA expression levels of SQSTM1 and ATG5 analyzed and quantified using RT-qPCR after knockdown of XBP1s. Data are represented as the means ± SEM of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 vs. control group. #, P < 0.05; ##, P < 0.01; ###, P < 0.001 vs. other group.

Article Snippet: The medium was replaced with fresh medium and the following treatments were initiated: (1) rapamycin (50 nM; TargetMol, Boston, MA, USA) for 3 h, 6 h, 12 h, and 24 h; (2) pretreatment with TG (Thapsigargin; 50 nM; an ERS activator; TargetMol) or 4-PBA (4-Phenylbutyric acid; 1 mM, an ERS inhibitor; Abcam, Cambridge, UK) or SC79 (10 μM; a PI3K/AKT pathway activator; TargetMol) or LY294002 (10 μM, a PI3K/AKT pathway inhibitor; TargetMol) for 2 h, followed by rapamycin treatment.

Techniques: Activation Assay, Knockdown, Expressing, Western Blot, Quantitative RT-PCR, Control