Journal: Neural Regeneration Research
Article Title: Enhancing m 6 A modification in the motor cortex facilitates corticospinal tract remodeling after spinal cord injury
doi: 10.4103/NRR.NRR-D-23-01477
Figure Lengend Snippet: METTL14 promotes axon regeneration by activating the Trib2/MAPK pathway. (A) Western blots of TRIB2, p-JNK and p-P38 proteins in NC- Mettl14 , sh- Mettl14 , and sh- Mettl14 +OE- Trib2 treated neurons. (B–D) Quantification of TRIB2, p-JNK and p-P38 protein expression levels in A ( n = 3 per group). (E–L) Relative expression of Ntrk3 , Cntf , Igf1r , NeuroD1 , Nrg1 , Pten , Cers2 , and Ptprs in NC- Mettl14 , sh- Mettl14 , and sh- Mettl14 +OE- Trib2 treated neurons by qPCR ( n = 3 per group). (M) Representative images of immunofluorescence staining of TUJ1 (red, AF594) in NC- Mettl14 , sh- Mettl14 , and sh- Mettl14 +OE- Trib2 treated neurons, showing that overexpression of TRIB2 counteracted the impact of Mettl14 knockdown. Scale bar: 100 μm. (N) Quantification of neurite length in N ( n = 4 per group). (O) Representative immunofluorescence images of the regenerative axons (TUJ1, green, AF488) on the microfluidic cultures, showing that overexpression of TRIB2 counteracted the impact of Mettl14 knockdown. Scale bar: 200 μm. (P) Quantification of total axonal length in G ( n = 4 per group). The data are presented as means ± SD. * P < 0.05, ** P < 0.01 (one-way analysis of variance followed by Tukey’s post hoc test (B–L, N, P)). Cers2: Ceramide synthase 2; Cntf: ciliary neurotrophic factor; DAPI: 4′,6-diamidino2-phenylindole; Igf1r: insulin like growth factor 1 receptor; JNK: Jun N terminal kinase; MAPK: mitogen-activated protein kinase; Mettl14: methyltransferase-like protein 14; NC: negative control; NeuroD1: neurogenic differentiation 1; Nrg1: neuregulin 1; Ntrk3: neurotrophic receptor tyrosine kinase 3; OE: over-expression; Pten: phosphatase and tensin homolog; Ptprs: protein tyrosine phosphatase receptor type S; Sh: short hairpin; Trib2: tribbles pseudokinase 2.
Article Snippet: P-P38 , Rabbit , WB: 1:750 , Wanleibio, Shenyang, China , WLP1576 , AB 2922420.
Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Over Expression, Knockdown, Negative Control