Journal: Neural Regeneration Research

Article Title: Salsolinol as an RNA m 6 A methylation inducer mediates dopaminergic neuronal death by regulating YAP1 and autophagy

doi: 10.4103/NRR.NRR-D-23-01592

Figure Lengend Snippet: Autophagy regulated by Sal is partly related to the inhibition of YAP1. (A) Sal treatment decreased the expression of p62 and increased LC3. (B) Autophagy-related proteins were examined in cells treated with Sal alone or in combination with BafA1 treatment. (C) In PC12 cells with YAP1 knockdown, the levels of p62 decreased and LC3 increased. Data are expressed as the mean ± SD ( n = 3/group). * P < 0.05, *** P < 0.001, vs. NC group (Student’s t -test). (D) In PC12 cells treated with Verteporfin, the levels of p62 decreased and LC3 increased. (E) A schematic model in which Sal acts as an RNA m 6 A methylation inducer, mediating dopaminergic neuronal death by regulating YAP1 and autophagy. ALKBH5: alkB homolog 5; BafA1: Bafilomycin A1; FTO: fat mass and obesity-associated protein; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; LC3: light chain 3; m 6 A: N 6 -methyladenosine; METTL3: methyltransferase-like 3; NC: normal control; p62: Sequestosome 1; Sal: salsolinol; siYAP1: small interfering RNA of YAP1; YAP1: Yes-associated protein 1; YTHDF2: YTH domain-containing family protein 2.

Article Snippet: The membrane was blocked with 5% skimmed milk and incubated with mouse anti-FTO monoclonal antibody (1:1000, Abcam, Cat# ab92821, RRID: AB_10565042), rabbit anti-ALKBH5 monoclonal antibody (1:1000, Abcam, Cat# ab195377, RRID: AB_2827986), rabbit anti-METTL3 monoclonal antibody (1:1000, Abcam, Cat# ab195352, RRID: AB_2721254), mouse anti-YAP1 monoclonal antibody (1:8000, Proteintech, Chicago, IL, USA, Cat# 66900-1-Ig, RRID: AB_2882229), mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) monoclonal antibody (1:100,000, Proteintech, Cat# 60004-1-Ig, RRID: AB_2107436), rabbit anti-YTH domain-containing family protein 1 (YTHDF1) polyclonal antibody (1:2000, Proteintech, Cat# 17479-1-AP, RRID: AB_2217473), rabbit anti-YTH domain-containing family protein 2 (YTHDF2) polyclonal antibody (1:5000, Proteintech, Cat# 24744-1-AP, RRID: AB_2687435), mouse anti-YTH domain-containing family protein 3 (YTHDF3) monoclonal antibody (1:1000, Santa Cruz Biotechnology, Texas, CA, USA, Cat# sc-377119, RRID: AB_2687436), rabbit anti-p62 polyclonal antibody (1:1000, MBL, Tokyo, Japan, Cat# PM045, RRID: AB_1279301), rabbit anti-light chain 3 beta (LC3B) polyclonal antibody (1:1000, Cell Signaling Technology, Boston, MA, USA, Cat# 2775, RRID:AB_915950), and rabbit anti-phospho-YAP1 (Ser127) (1:1000, Cell Signaling Technology, Cat# 13008, RRID: AB_2650553) at 4°C overnight.

Techniques: Inhibition, Expressing, Knockdown, Methylation, Control, Small Interfering RNA