Fig. 3

Blocking increased spinal Peli1 reverses CCI-induced microglial activation and the production of pro-inflammatory cytokines. a, b Timeline schematic of experimental paradigm. c Representative images of Iba1 immunofluorescence (red) on the ipsilateral side of dorsal horn with a high magnification image (boxed, scale bar 50 μm). d Qualitative data showing the inhibitory effect of Peli1 shRNA on the number of Iba1⁺ cells in the dorsal horn (n = 4). One-way ANOVA (F_3,12 = 28.6). e, f Administration of Peli1 shRNA inhibits CCI-induced upregulation of the microglial marker Iba1 (e, n = 5–6) and CD11b (f, n = 5–6) mRNA expression in L4-L5 ipsilateral dorsal spinal cord. One-way ANOVA [(e) F_3,17 = 28.3; (f) F_3,15 = 18.2)]. g, h Representative images and quantitation of Peli1 immunofluorescence showing the inhibitory effect of Peli1 siRNA on established microglial activation on the ipsilateral side of the dorsal horn (scale bar = 50 μm, n = 4). One-way ANOVA (F_3,12 = 62.7). i–k The levels of TNF-α (i, n = 4–5), IL-6 (j, n = 4–5), and IL-1β (k, n = 4–5) were measured on day 7 by ELISA. Spinal Peli1 knockdown inhibits CCI-induced TNF-α, IL-6, and IL-1β production in the ipsilateral dorsal horn. One-way ANOVA [(i) F_5,19 = 10.9; (j) F_5,18 = 11.9; (k) F_5,17 = 13.9]. Results are expressed as the Mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 compared with indicated group