TXNIP Antibody - #DF7506

Fig. 8. TXNIP may be mainly derived from microglia. (A) Co-localization of TXNIP with neurons, microglia, and astrocytes in the hippocampus (Scale bar = 20 μm). (B) Fluorescence co-localization distribution curves of TXNIP with NeuN, Iba-1, and GFAP, respectively. (C) Microglial depletion efficiency in mouse brain by PLX5622 treatment, showing Iba-1+ microglia (green) and DAPI+ nuclei (blue). Scale bar = 50 μm. (D) Quantitative analysis of microglial cell numbers in the hippocampus. (E) Quantitative analysis of microglial cell numbers in the cortex. (F) Effect of PLX5622-mediated microglial depletion on TXNIP level (red) in I/R mouse brain, with DAPI + nuclei (blue). Scale bar = 50 μm. (G) Analysis of TXNIP mean fluorescence intensity in II/R mouse brain. (Data presented as mean ± SEM; ∗∗P < 0.01, ∗∗∗P < 0.001 vs. control group).

Fig. 6 Identification and validation of TXNIP as a target gene for miR-15b-5p. (A, B) miRWalk analysis and Gene Ontology (GO) analysis identified 25 genes in the intersection set of Cellular Component (CC), Biological Process (BP), Reaction, Molecular Function (MF), and KEGG pathways. (C, D) Protein-Protein Interaction (PPI) network and KEGG pathway analysis of the 25 identified genes, revealing associations with ROS signal transduction, NLRP3 inflammasome activation, and inflammation, all closely related to BMSCs osteogenic differentiation. (E) Wild-type (WT) and mutated miR-15b-5p recognition sites within the TXNIP 3’ UTR were generated for the dual-luciferase reporter assay. (F) Luciferase reporter assay demonstrating decreased luciferase activity of WT-Txnip in response to miR-15b-5p overexpression (n = 3 per group). (G, H) Western blotting analysis of Txnip protein levels in BMSCs, indicating upregulation in response to miR-15b-5p inhibition (n = 3 per group). The error bars represent the means ± SDs.

Fig. 10. miR-17-5p shuttling by MSCs-Exo inhibited Kupffer cell pyroptosis by regulating the TXNIP-NLRP3-Caspase1 signaling pathway. (A) Representative immunofluorescence staining images of TXNIP (green) in Kupffer cells treated with MSCs-ExomiR−Ctrl−KD and MSCs-ExomiR−17−5p KD under 300µM H2O2 stimulation. (B) Comparison of the protein levels of TXNIP, NLRP3 and Cle-caspase1 in the Control, H2O2, H2O2 + MSCs-ExomiR−Ctrl−KD and H2O2 + MSCs-ExomiR−17−5p KD groups. (C) The TXNIP protein was overexpressed in the OE-TXNIP Kupffer cells. The protein levels of TXNIP, NLRP3 and Cle-caspase1 were measured in the H2O2, H2O2 + OE-TXNIP, H2O2 + OE-TXNIP + MSCs-ExomiR−Ctrl−KD and H2O2 + OE-TXNIP + MSCs-ExomiR−17−5p KD groups

Figure 1 Induction of TRX and Nrf2 by GGA in the ICD model. (A,B) Immunohistochemistry was performed to detect the effects of GGA on TRX induction and Nrf2 activation. GGA induces the production of TRX, especially after oral administration. Normal = mice without any treatment. NS = normal saline; bars 100 μm. The most pronounced nuclear translocation of Nrf2 was in the GGA gavage-treated group, followed by the GGA topically applied group; bars 50 μm; n = 5 mice for each group. Cells with nuclear translocation are indicated by arrows. (C,D) The mean of IOD expression of TRX and Nrf2. Values are shown as the mean ± SD (* p < 0.05, ** p < 0.01, **** p < 0.0001; one-way ANOVA with post hoc Tukey’s).

Figure 9. Effect of SMS on TXNIP/NLRP3 pathway in FFA induced HepG2 cells. Cells were treated with different concentrations of SMS and then induced with FFA (1 mM) for 24 h each. (A–C) The fluorescence intensity of TXNIP, NLRP3 and GSDMD was detected by immunofluorescence staining. The green color represented TXNIP, NLRP3 and GSDMD staining. (D) The levels of TXNIP mRNA expression were detected by qRT-PCR. (E) The expression of TXNIP, NLRP3, caspase 1, cleaved-caspase 1, GSDMD and GSDMD-N were detected by Western blotting, and their expression was normalized relative to β-actin. Results are mean ± SD (n = 3). #P < 0.05, ##P < 0.01 vs. CON group; *P < 0.05, **P < 0.01 vs. FFA group.

Figure 10. Effect of SMS on the inflammation in FFA induced TXNIP knockdown HepG2 cells. (A) The protein level of TXNIP were detected by Western blotting with siRNA-TXNIP. (B) Cells were treated with SMS (100 μg/mL) and then induced with FFA (1 mM) for 24 h each. The levels of TXNIP mRNA expression were determined by qRT-PCR. The levels of (C) LDH, (D) IL-1β, (E) IL-18, (F) TC and (G) TG were detected by reagent kits. Results are mean ± SD (n = 3). #P < 0.05, ##P < 0.01 vs. siR-Control group; §P < 0.05, §§P < 0.01 vs. CON group; *P < 0.05, **P < 0.01 vs. FFA group.