Fig. 8: MAF1 mediates the regulatory effect of PCIF1 on neuropathic pain and comorbid anxiety.
a Reduced S1HL MAF1 protein 21 days post AAV-DIO-shMaf1 injection in CaMK2α-Cre mice (n = 8 mice). Knockdown of S1HL Maf1 or Co-injection of CRISPR-dCasRx-Pcif1 + gRNA-26/60 induced contralateral mechanical allodynia (b, c, e, f) and thermal hyperalgesia (d, g) (n = 10 mice). h MAF1 levels increased after injection of AAV-DIO-MAF1 into CaMK2α-Cre mice (n = 12 mice). i–k Overexpressing MAF1 in contralateral S1HL attenuated SNI-induced mechanical allodynia (i, j) and thermal hyperalgesia (k) (n = 10 mice). l S1HL PCIF1/MAF1 levels after co-injection of AAV-DIO-Pcif1 ± AAV-DIO-Maf1 in naïve CaMK2α-Cre mice (n = 8 mice). m–o MAF1 overexpression blocked PCIF1-induced pain-like behaviors (n = 10 mice). p Schematic (left, drawn by figdraw.com) and GCaMP6s expression image (right) for fiber photometry. Heatmaps (q), average Ca2+ transients (r), and AUC quantification (s) of GCaMP6s signals recorded from S1HLGlu neurons as mice received 0.4 g von Frey stimulation (n = 24 trails from 8 mice). Scale bar, 200 µm. t Schematic of patch-clamp recordings from labeled S1HLGlu neurons in slices. Sample traces (u), statistical data (v), and rheobase of the spike (w) for action potential firing and the membrane potential (x) recorded from S1HLGlu neurons (n = 13 cells from 4 mice). Data are represented as mean ± SEM. S1 samples from two mice were pooled to create one sample (a, h and l). Student’s unpaired t-test (two-tailed) was used for (a) and (v). Two-way ANOVA followed with Bonferroni’s post hoc test was used for (b–d). Two-way ANOVA followed with Tukey’s post hoc test was used for (e–g). One-way ANOVA followed with Tukey’s post hoc test was used for (h, l, s, w and x). Two-way ANOVA followed with Tukey’s post hoc test was used for panels (i–k and m–o).
