Fig. 7: Increased PCIF1 leads to SNI-induced downregulation of MAF1 in S1HL. | Nature Communications

Fig. 7: Increased PCIF1 leads to SNI-induced downregulation of MAF1 in S1HL.

From: SERBP1-PCIF1 complex-controlled m6Am modification in glutamatergic neurons of the primary somatosensory cortex is required for neuropathic pain in mice

Fig. 7

a MAF1 protein levels on day 21 after injection of AAV-DIO-Pcif1 (AAV-Pcif1; to induce PCIF1 expression) or AAV-DIO-mCherry (DIO-mCherry) into S1HL of naïve CaMK2α-Cre mice (n = 10 mice). b The level of MAF1 protein in SNI or Sham Pcif1fl/fl mice preinjected with AAV-CaMK2α-Cre (CaMK2α-Cre; to knock out PCIF1) or AAV-CaMK2α-mCherry (CaMK2α-mCherry) into S1HL. Tissue was harvested 21 days after injection (n = 12 mice). c, d Co-expression analysis of MAF1 (red) with NeuN (a neuronal marker, cyan), S100β (an astrocyte marker, cyan), or Iba1 (a microglia marker, cyan) immunofluorescence staining in the S1HL of naïve mice. n = 4 mice. Scale bar, 200 μm. e Co-expression analysis of Maf1 with Serbp1 and Pcif1 in CaMK2α neurons by single-cell PCR. Numbers 1-8 represent eight individual neurons. M, DNA marker. f CRISPR-dCasRx/PCIF1 “writing” m6Am to the given site in Maf1 mRNA. gRNA, small guide RNA. g Identification of dCasRx-Pcif1 fusion protein expression on day 5 after microinjection of CRISPR-dCasRx-Pcif1 into S1HL. h, i The Maf1 m6Am (h) and MAF1 protein (i) levels on day 5 after co-microinjection of CRISPR-dCasRx-Pcif1 and gRNA-26 or gRNA-60 into S1HL in naïve mice (n = 10 mice). For (a, b and h, i), data are represented as mean ± SEM. S1 samples from two mice were pooled to create one sample (a, b, h and i). Student’s unpaired t-test (two-tailed) was used for (a). One-way ANOVA followed with Tukey’s post hoc test was used for (b, h and i).

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