Journal: Cell reports

Article Title: Feedforward miR-181d degradation modulates population variance of methyl-guanine methyl transferase and temozolomide resistance

doi: 10.1016/j.celrep.2025.116516

Figure Lengend Snippet: PNPT1 expression analysis in clinical glioblastoma specimens. RNA ( n = 5) and protein ( n = 3) extracted from matched adjacent brain tissue specimens were analyzed for PNPT1. (A) PNPT1 mRNA expression by RT-qPCR. Statistical significance: * p < 0.05 and *** p < 0.001. (B) PNPT1 protein expression by western blotting. (C) Left: PNPT1 protein structure labeled with numbers represents the specific position for the amino acid substitution mutation. Right table: summary of amino acid substitution within PNPT1 domains. Bottom: CMK3 WT and PNPT1-KO clones (PNPT1-KO1 and PNPT1-KO2) were transfected with either WT or mutated PNPT1 constructs. miR-181d expression was quantified by RT-qPCR. (D) In vitro miR-181d degradation assay by immunoprecipitated WT or mutant PNPT1. GFP-tagged PNPT1 protein was immunoprecipitated using an anti-GFP antibody from WT or D538A mutant PNPT1-expressing lines and incubated with miR-181d. RNA was extracted and quantified by RT-qPCR. Data are represented as mean ± SD.

Article Snippet: Anti-GFP antibody , OriGene , Cat# TA150039; RRID: AB_2622255.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Labeling, Mutagenesis, Clone Assay, Transfection, Construct, In Vitro, Degradation Assay, Immunoprecipitation, Incubation