Fig. 3: Upregulating S1HL^Glu PCIF1 induces neuropathic pain- like behavior.

a Timeline for virus injection and behavior tests in CaMK2α-Cre mice. b Schematic of the experimental paradigm (left) and a representative image showing mCherry expression in S1HL after injection of AAV-DIO-Pcif1 or AAV-DIO-mCherry into CaMK2α-Cre mice (right). Scale bar, 500 µm (left). PCIF1 (c) and m⁶Am (d) levels significantly increased 28 days post AAV-DIO-Pcif1 (vs. mCherry) in CaMK2α-Cre mice (n = 8 mice). PCIF1 upregulation in S1HL^Glu neurons induced contralateral mechanical allodynia (e, f), thermal hyperalgesia (g), as measured by conditioned place preference (h) (e–g: n = 8 mice, h: n = 7 mice). i Schematic (drawn by figdraw.com) and image of GCaMP6s recording setup in awake mice (scale bar: 500 µm). Scale bar, 500 µm. Heatmaps (j), average Ca²⁺ transients (k), and AUC quantification of the GCaMP6s signal (l) from S1HL^Glu neurons of CaMK2α-Cre mice receiving 0.4 g von Frey stimulation (n = 24 trails from 8 mice). m Patch-clamp schematic in S1HL^Glu neurons. Sample traces (n), statistical data (o), and rheobase of the spike (p) for action potential firings and the membrane potential (q) recorded from S1HL^Glu neurons in mice with upregulated PCIF1 expression, as described for (c, d) above (DIO-mCherry group: n = 14 cells from 4 mice, DIO-Pcif1 group: n = 13 cells from 4 mice). The data are represented as mean ± SEM. S1 samples from two mice were pooled to create one sample (c, d). Student’s unpaired t-test (two-tailed) was used for (c, d, h, p and q). Two-way ANOVA followed with Bonferroni’s post hoc test was used for (e–g and o). One-way ANOVA followed with Bonferroni’s post hoc test was used for (l).