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. 2020 Jun 28;105(9):2983–2995. doi: 10.1210/clinem/dgaa413

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Figure 3. — A, Time-course of peroxisome-proliferator-activated receptor β/δ (PPARβ/δ) agonist GW0742 (GW) effects on endometrial stromal cell (ESC) protein lysates by Western blotting. GW treatment for 15 minutes to 48 hours showed changes in nuclear receptors: PPARβ/δ (49 kDa, panel 1), estrogen receptor α (ERα) (66 kDa, panel 2), PR-A and -B (90 and 116 kDa, respectively, panel 3). Phosphoproteins of the extracellularly regulated kinase (ERK)1/2 cascade included phospho-MEK1/2 (45 kDa, panel 4), phospho-ERK1/2 (42 kDa, panel 5), and phospho-p90 RSK (90 kDa, panel 6). Connexin 43 (Cx43) isoforms: phospho-Cx43 Ser367 (43 kDa, panel 7), nonphospho-Cx43 Ser368 (43 kDa, panel 8). β-Actin levels (42 kDa, panel 9) are shown. Lane 1 represents untreated control cells and lane 9, ESCs treated for 24 hours with all-trans retinoic acid (RA) as a positive control. B, Under the same conditions, ESC supernatant concentrations of vascular endothelial growth factor (VEGF) were determined by enzyme-linked immunosorbent assay. Means ± SEM of triplicate samples are shown. GW at 24 and 48 hours and RA at 24 hours each significantly stimulated VEGF secretion over control conditions (P < .05, t tests with Bonferroni corrections, n = 3). C, Time-course effects of PPARβ/δ antagonist, GSK0660 (GSK) on ESC protein markers by Western blotting. Phosphoproteins of the extracellularly regulated kinase (ERK)1/2 cascade included phospho-MEK1/2 (45 kDa, panel 1), phospho-ERK1/2 (42 kDa, panel 2), phospho-p90 RSK (90 kDa, panel 3), Cx43 isoforms: Phospho-Cx43 Ser367 (43 kDa, panel 4), phospho-Cx43 Ser368 (43 kDa, panel 5), and nonphospho-Cx43 Ser368 (43 kDa, panel 6). β-Actin levels (42 kDa, panel 7) are shown. Lane 1 represents untreated control cells.