Figure 2.

Efficacy of AAV-KLF7 gene transfer to HT22 cells following stretch and OGD in vitro.

(A) Representative western blot analysis of KLF7 expression in cultured HT22 cells at 1 day after stretch and OGD in vitro. (B) Quantitative results of the relative expression of KLF7 protein detected by western blot. The relative expression is expressed as the optical density ratio to GAPDH. (C) qRT-PCR analysis of KLF7 mRNA expression in the cultured HT22 cells at 1 day after stretch and OGD. (D) Immunofluorescence staining of KLF7 (green, stained by fluorescein isothiocyanate) and βIII-tubulin (red, stained by Alexa 555) with nuclear staining (DAPI; blue) in the three groups. The merged image shows that HT22 cells were co-labeled by KLF7, βIII-tubulin, and DAPI. The expression of KLF7 was increased after AAV-KLF7 infection compared with the other groups, and β-III-tubulin expression was also elevated in AAV-KLF7 cells compared with AAV-NC cells. Scale bars: 100 μm. (E, F) Quantitative results of the immunopositivity of KLF7 (E) and βIII-tubulin (F). Data are expressed as the mean ± SD. Each experiment was repeated three (A–C) or six (D–F) times. *P < 0.05, **P < 0.01, ***P < 0.001 (one-way analysis of variance followed by Tukey’s post hoc test). AAV: Adeno-associated virus; DAPI: 4′,6-diamidino-2-phenylindole; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; KLF7: Krüppel-like factor 7; NC: negative control; OGD: oxygen-glucose deprivation; qRT-PCR: quantitative real-time polymerase chain reaction.