Fig. 2

LncARAT upregulates CCL2 expression in SCs and promotes H3K4me3 at the CCL2 promoter by interacting with KMT2A. A Western blot and quantitative analysis of CCL2 expression in injured nerves 4 dpi (n = 5). B IF assay of CCL2 (green) and S100β (red) in injured nerves 4 dpi after lncARAT overexpression or knockdown. Scale bar, 50 µm. C Western blot and quantitative analysis of CCL2 expression in primary SCs after lncARAT overexpression for 48 h (n = 3). D Sequential deletions of the CCL2 promoter linked to Renilla luciferase were constructed and then transfected into SCs to assess their transcriptional activity 48 h later. E Left panel, schematic presentation of the potential lncARAT binding sites in the CCL2 promoter. Right panel, ChIRP assessment of lncARAT-associated chromatin in SCs. F Genomic neighborhood of CCL2. Genome browser tracks from the UCSC genome browser showing H3K4me3 occupancy near CCL2. G IF assay of H3K4me3 levels (green) and S100β levels (red) in injured nerves 4 dpi after lncARAT knockdown. Scale bar, 50 µm. H RNA pulldown followed by western blot analysis of KMT2A, KMT2B, and KMT2D in SCs. I RIP assay using anti-KMT2A, KMT2B, or KMT2D antibodies in SCs. U1 was used as the negative control. ChIP analyses using KMT2A antibody (J) and H3K4me3 antibody (K) on the regulatory regions of CCL2 genes in SCs 72 h after lncARAT overexpression. *p < 0.05. **p < 0.01