Figure 5.

The JAK2/STAT3 inhibitor AG490 reverses the protective effects of KLF7 on stretch-and OGD-induced cell apoptosis.

(A) Bcl-2, Bax, and C-caspase3 expression of cultured HT22 cells at 1 day in vitro. (B–D) Quantitative results of the relative expression of Bcl-2, Bax, and C-caspase3. The relative expression is expressed as the optical density ratio to GAPDH. (E) HT22 cell apoptosis measured by PI staining, with nuclear staining (Hoechst 33342; blue). AAV-KLF7 treatment reduced the expression of C-caspase-3 and Bax and elevated the expression of Bcl-2 compared with AAV-NC treatment. However, the effects of KLF7 on C-caspase-3, Bax, and Bcl-2 were abrogated by AG490 treatment. Scale bars: 100 μm. (F) Quantitative results of the percentage of PI-positive cells. (G) LDH activity was measured using ELISA. (H) Caspase-3 activity was detected using ELISA. (I) Cell viability was measured using CCK-8. Data are expressed as the mean ± SD. Each experiment was repeated three (A–D) or six (E–I) times. *P < 0.05, vs. control group; #P < 0.05, vs. stretch + OGD + AAV-NC group; &P < 0.05, vs. stretch + OGD + AAV-KLF7 group (one-way analysis of variance followed by Tukey’s post hoc test). AAV: Adeno-associated virus; Bax: Bcl-2-associated X protein; Bcl-2: B-cell lymphoma 2; C-caspase3: cleaved caspase-3; CCK-8: Cell Counting Kit-8; DAPI: 4′,6-diamidino-2-phenylindole; ELISA: enzyme-linked immunosorbent assay; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; KLF7: Krüppel-like factor 7; LDH: lactate dehydrogenase; NC: negative control; OGD: oxygen–glucose deprivation; PI: propidium iodide.