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. Author manuscript; available in PMC: 2026 Jan 6.
Published in final edited form as: Cell Rep. 2025 Nov 10;44(11):116516. doi: 10.1016/j.celrep.2025.116516

Figure 3. Ribonuclease activity of PNPT1-dependent degradation of miR-181d.

Figure 3.

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.