Fig. 2: m6A modification of the lncRNA NORAD contributes to NPC senescence.
From: WTAP-mediated m6A modification of lncRNA NORAD promotes intervertebral disc degeneration
a, b PEAK and gene number analysis of Me-RIP-Seq results of NPCs. c, d PEAK distribution of gene regions of m6A-modified transcripts. e Representative motif of m6A modification of transcripts in normal and senescent NPCs, two-sided Fisher’s exact test with adjustments for multiple testing. f Correlation analysis of m6A and transcript-differ genes. g m6A level of NORAD transcripts in normal and senescent NPCs by Me-RIP-qPCR. Data are shown as the mean ± SD, n = 3, P = 0.0007, *P < 0.05, two-tailed unpaired Student’s t test. h, i Expression of NORAD in NPCs (n = 3, P = 0.0094) and NP tissues (n = 58, P < 0.0001) by RT-qPCR. Data are shown as the mean ± SD, **P < 0.01, ****P < 0.0001, two-tailed unpaired Student’s t test. j, k NORAD expression in NPCs and NP tissues by FISH (red: NORAD; blue: DAPI; scale bar: 20 μm; 50 μm). l Radiographic presentation of control and IVDD model by MRI, μCT and X-Ray. m Histological of IVDD model analyzed by HE staining and IF analysis (green: Collagen II; red: P16; blue: DAPI; scale bar: 100 μm; 25 μm). n Radiographic analysis of IVDD model by DHI% in control (P = 0.0005) and puncture group (P < 0.0001); n = 6, ***P < 0.001, ****P < 0.0001. Histological score of IVDD model in control (P = 0.0024) and puncture group (P < 0.0001); two-tailed unpaired Student’s t test. n = 6, **P < 0.01, ****P < 0.0001, two-tailed unpaired Student’s t test. o NORAD expression in NPCs from WT and KO mice by FISH (red: NORAD; blue: DAPI; scale bar: 20 μm). p, q Protein level of ACAN (P = 0.0022), Collagen II (P < 0.0001), P16 (P < 0.0001), P21 (P = 0.0005) in NPCs from WT and KO mice by western blot, GAPDH was used as a loading control. Data are shown as the mean ± SD, n = 7. **P < 0.01, ***P < 0.001, ****P < 0.0001, two-tailed unpaired Student’s t test. r Protein level of P16 and P21 in NPCs with knockdown or overexpression of NORAD (sh-ND: shRNA of NORAD; OE-ND: overexpression of NORAD); GAPDH was used as a loading control. s SA-β-gal staining of NPCs with knockdown or overexpression of NORAD (Scale bar: 200 μm).
