Fig. 4. Reduction of apoptosis and autophagic stress by treatment with silibinin in the KA-treated hippocampus. (A, B) Double-immunofluorescence staining.

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Fig. 4. Reduction of apoptosis and autophagic stress by treatment with silibinin in the KA-treated hippocampus. (A, B) Double-immunofluorescence staining for NeuN (green) and c-caspase-3 (red), and NeuN (green) and c-PARP-1 (red) in the hippocampal CA1 region 2 days after KA treatment. Insets delimited by the solid line in the panels on the right show high magnification portions of the image. Scale bars are given for both the main image (40x) and insets (100x): 20 µm and 10 µm. (C, D) Western blot analysis of c-caspase-3, caspase-3, c-PARP-1, and PARP-1 expression in the hippocampus 2 days after KA treatment. The density of c-caspase-3, caspase-3, c-PARP-1, and PARP-1 bands was normalized to the β-actin band for each sample. *p<0.001 and **p<0.01 vs. PBS alone, #p<0.01 vs. KA alone (one-way ANOVA and Tukey's post-hoc analysis; n=4, each experimental group). All values are expressed as the mean±SEM. (E) Double-immunofluorescence staining for NeuN (green) and LC3B (red) in the hippocampal CA1 region 2 days after KA treatment. Scale bars are given for both the main image (40x) and insets (100x): 20 µm and 10 µm. (F, G) Western blot analysis of LC3 in the hippocampus 2 days after KA treatment. The density of LC3-II band was normalized to the β-actin band for each sample. *p<0.001 vs. PBS alone, #p<0.001 vs. KA alone (one-way ANOVA and Tukey's post-hoc analysis; n=4, each experimental group). All values are expressed as the mean±SEM. Fig. 4. Reduction of apoptosis and autophagic stress by treatment with silibinin in the KA-treated hippocampus. (A, B) Double- immunofluorescence staining for NeuN (green) and c-caspase-3 (red), and NeuN (green) and c-PARP-1 (red) in the hippocampal CA1 region 2 days after KA treatment. Insets delimited by the solid line in the panels… Exp Neurobiol. 2017 Oct;26(5):266-277. https://doi.org/10.5607/en.2017.26.5.266