Fig. 6
Spinal Peli1 is necessary for MAPK/NF-κB activation and TRAF6 ubiquitination after CCI. a–c Peli1 shRNA was given 3 days prior to the beginning of CCI. L4-L5 spinal cord tissues were collected at 7 days after CCI. Quantitative western blot analysis showing inhibitory effects of Peli1 shRNA on CCI-induced increased phosphorylation of ERK (a, n = 4–6), p38 (b, n = 4–5), and JNK (c, n = 4). One-way ANOVA [(a) F5,21 = 12.3; (b) F5,20 = 24.9; (c) F5,18 = 14.5]. d, e Double immunofluorescence staining showing lower colocalization of p-ERK (red) and p-p38 (green) with microglia marker Iba1 (red or green) by Peli1 shRNA in the ipsilateral dorsal horn at 7 days (scale bar 50 μm). f, g Peli1 shRNA suppresses CCI-induced increases in p-NF-κB p65 (f, n = 4–5) and P-Akt (g, n = 4–5) in the ipsilateral spinal cord on day 7 after injury. One-way ANOVA [(f) F5,19 = 42.8; (g) F5,19 = 10.2]. h Analysis of K63-linked ubiquitination of TRAF6 in the ipsilateral spinal cord that transfected with Peli1 shRNA or scrambled shRNA 3 days before the beginning of CCI. Tissues were collected at 7 days after CCI (n = 4). Results are expressed as the Mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 compared with indicated group