Fig 2: Downregulation of miR-17 impaired glucose uptake by promoting oxidative stress via targeting keap1. A-D. The representative western blot images (A) and quantification (B-D) of genes related to oxidative stress. Gapdh was used as the loading control. C2C12 myotubes were transfected with 50 nM agomiR-17 or 200 nM antagomir-17 for 48 h (n = 3 independent experiments). E. The miR-17 target regions in 3'UTR of Keap1. Mmu, mouse; Rno, rat; Hsa, human. WT: a truncated Keap1-3'UTR with wild-type miR-17 binding site; MT: a truncated Keap1-3'UTR with mutated miR-17 binding site. F-G. The relative luciferase activity in C2C12 myoblasts transfected with reporter vector containing the wild-type (F) or mutated (G) Keap1 3'UTR together with agomiR-17, antagomiR-17 or corresponding controls (n = 4 independent experiments). H-J. The representative western blot images (H) and quantification (I-J) of pAkt and mGlut4 in C2C12 myotubes treated with antagomiR-17 (n = 3 independent experiments). Gapdh was used as the loading control. K. The insulin-stimulated 2-DG uptake in C2C12 myotubes treated with antagomiR-17 (n = 3 independent experiments). L. The expression of miR-200a in C2C12 myotubes transfected with agomiR-17 and antagomiR-17 (n = 3 independent experiments). U6 was used to normalized miR-200a expression. M-P. C2C12 myotubes were transfected with 50 nM agomiR-17 and treated with 25 µM DEHP (n = 3 independent experiments). M-N. The mRNA expression of Keap1 and Nrf2. Gapdh was used as the loading control. O. The calculated GSH/GSSG ratio. The levels of GSH and GSSG were shown in Figure S8C-D. P. The insulin-stimulated 2-DG uptake. Q-S. C2C12 myotubes were co-treated with 25 µM DEHP, antagomir-17, SFN or corresponding controls (n = 3 independent experiments). Q. The expression of miR-200a normalized by U6. R. The mRNA expression of Txnip. Gapdh was used as the loading control. S. The insulin-stimulated 2-DG uptake. All data were presented as the mean ± SEM. *P < 0.05, **P < 0.01 vs. corresponding control as indicated.

Fig 3: Neuroprotective effects of miR-25-5p on CTX TNA2 cells in response to LPS stimulation. (A and B) Effects of miR-25-5p mimic on miR-25-5p and TXNIP mRNA (A), TXNIP protein fold changes (B) of LPS-stimulated CTX TNA2 cells. (C) Cell viability of CTX TNA2 cells in response to 0 µg/mL, 1 µg/mL, 2 µg/mL, 5 µg/mL, and 10 µg/mL LPS stimulation at 12 h. (D–G) Effects of miR-25-5p mimic on cell viability (D), cleaved caspase-1 and NLRP3 protein fold changes determined by western blot (E), relative MFI of DCF (F), and percentage of apoptosis (G) of LPS-stimulated CTX TNA2 cells. The relative expression levels of miRNA and mRNA were determined by qPCR and protein levels were determined by Western blot. The independent t-test was applied to the comparison between groups (A, B, D–G). One-way analysis of variance (ANOVA), followed by Tukey’s test was used to identify the significant differences among multiple groups (C). Values are shown as the mean ± SD. Con: negative control of LPS, LPS: lipopolysaccharide, mimic NC: negative control of miR-25-5p mimic, PI: propidium iodide, MFI: mean fluorescence intensity. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001.

Fig 4: Mutation and modification feature of TXNIP in different TCGA tumors. A The genetic alteration type and frequency of TXNIP in various cancers. B The three-dimensional structure of TXNIP protein. C Mutation site of the TXNIP. D Correlation analysis of tumor mutational burden (TMB) with TXNIP gene expression. E Correlation analysis of microsatellite instability (MSI) with TXNIP gene expression. F Correlation analysis of four DNA methyltransferases (DNMT1, DNMT2, DNMT3A, DNMT3B) with TXNIP gene expression. G Correlation analysis of five MMR genes (EPCAM, MLH1, MSH2, MSH6 and PMS2) with TXNIP gene expression. H The predicted ubiquitination and phosphorylation sites of TXNIP protein

Fig 5: Correlation of TXNIP expression with immune and matrix scores, immune checkpoints, and neoantigens in different tumors of TCGA. A–C Correlation of TXNIP expression with the immune and matrix scores in LUAD, LUSC, COAD, and STAD. D Correlation of TXNIP expression with 47 transcripts related to immune checkpoints. E–K Correlation of TXNIP expression with the numbers of neoantigen