Fig. 3

miR-34b-5p promotes KA-induced astrocytes apoptosis by targeting Bcl-2. a Representative images of three repeated TUNEL staining experiments on cultured astrocytes. The astrocytes are pre-treated with of miR-34b-5p mimics, inhibitors, mimics negative control (mNC), and inhibitors negative control (i NC), followed by KA treatment; scale bar 50 μm. b Relative miR-34b-5p levels after different treatments; (one-way ANOVA, n = 5, F _4,20  = 866.9, ***P < 0.0001). c Percentage of TUNEL positive astrocytes over total counted astrocytes represented as apoptotic rate. Cells were treated with the four indicated treatments before fixing and performing TUNEL staining experiments (one-way ANOVA, n = 5, F _4,20 = 67.45, ***P < 0.0001). d Luciferase assay. Astrocytes was transfected with WT Bcl-2 3′-UTR or Mutant Bcl2 3′-UTR followed by treating either with control or miR-34b-5p mimics. Luciferase activity is indicated by the ratio of firefly luciferase activity to Renilla luciferase activity (Between miR-34b-5p mimics treated with WT-UTR or MUT-UTR, n = 3, t ₄ = 7.10, **P = 0.0021). e Graphic picture shows miR-34b-5p sequence match with rat Bcl-2 mRNA 3′-UTR. f Quantitative PCR shows Bcl-2 mRNA level in response to different treatments (one-way ANOVA, n = 5, F _4,20 = 0.4576, P = 0.7658)