Fig. 5

The impact of RN7SL1 knockdown on neuronal apoptosis and morphology through microglia. (a, b) Knockdown efficiency of smart silencer (ssi) directed against Rn7sl1 in rat primary microglia (a) and BV2 microglial cell lines (b), as determined by qRT‒PCR and normalized to ACTB expression. (c, d) Expression of proinflammatory cytokines in primary microglia (c) and BV2 cells (d) as determined by qRT‒PCR and normalized to ACTB expression. ssi-NC, cells treated with NC smart silencer as knockdown control; ssi-Rn7sl1, cells subjected to Rn7sl1 knockdown using Rn7sl1 smart silencer; ssi-NC + oAβ, knockdown control cells subjected to 5 µM oAβ stimulation for 24 h; ssi-NC + oAβ, knockdown control cells treated with 5 µM oAβ stimulation for 24 h; ssi-Rn7sl1 + oAβ, Rn7sl1-knockdown cells treated with 5 µM oAβ stimulation for 24 h. (e) Representative immunofluorescence images of BV2 cells phagocytosing latex beads in four groups and phagocytosis rate statistics. % Phagocytosis represents the number of BV2 cells that phagocytosed latex beads divided by total number of IBA1⁺ (microglial marker) cells (n = 10/group). (f) Flow cytometry histogram showing the phagocytosis rate of BV2 cells in four groups. % Phagocytosis represents the percentage of FITC⁺ cells (cells that phagocytosed fluorescent beads) within the gated total cell population (n = 6/group). (g) Representative images showing the effects of Rn7sl1 knockdown on BV2 cell migration. To quantify BV2 cell migration, absorbance at 570 nm was measured after crystal violet staining and washing (n = 6/group). Values represent mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, as determined by one-way ANOVA (for comparing multiple groups) or Student’s t-test (for comparing two groups)