Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: Knockout of GSDMD alleviated renal tubule morphological changes in OXO/adenine-induced mice and attenuated fibrosis (A and B) Histological changes in renal tubules after HE (A) and Masson (B) staining in different groups. Scale bar 50 μm and 100μm. (C and D) Western blot showing α-SMA levels in renal tissue from different groups (n = 6 animals per group). (C and E) Western blot showing E-cadherin levels in renal tissue from different groups (n = 5 animals per group). (F and G) RT‒PCR analysis of fibronectin (F) and collagen I (G) levels in renal tissue from different groups (n = 5 animals per group). (H) RT‒PCR analysis of IL-1β levels in renal tissue from different groups (n = 6 animals per group). Data for (D)-(H) are presented as mean ± SEM. One-way ANOVA with Student-Newman-Keuls test for (D–H). WT, wild type. Norm-diet, normal diet. UA, uric acid.

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: Knock-Out, Staining, Western Blot

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: The primer sequences used in RT‒PCR

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques:

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: GSDMD encouraged IL-1β expression and fibrosis in NRK-52E cells in vitro (A–D) HK-2 cells were stimulated with uric acid for 48 h. Western blot showing caspase-1 (A and B), GSDMD (A and C) and RIG-I (A and D) levels in HK-2 cells in the different groups (n = 3 per group). (E–H) NRK-52E cells were stimulated with uric acid for 48 h. Western blot showing cleaved-caspase-1 (E and F), caspase-1 (E and G) and RIG-I (E and H) levels in NRK-52E cells in the different group (n = 6 per group). (I–P) NRK-52E cells were transfected with GSDMD-siRNA or NC-siRNA and treated with UA for 48 h. Western blot showing GSDMD (I and J) and GSDMD-N (I and K) levels after UA treatment with or without GSDMD knockdown (n = 6 per group). (L) IL-1β level in culture medium (n = 6 per group). (M–P) Western blot showing α-SMA (M and N), E-cadherin (M and O) and fibronectin (M and P) levels after UA treatment with or without GSDMD knockdown (n = 6 per group). Data for (B–D), (F–H), (J–L) and (N–P) are presented as mean ± SEM. Student’s t test for (B–D) and (F–H). One-way ANOVA with Student-Newman-Keuls test for (J–L) and (N–P). UA, uric acid.

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: Expressing, In Vitro, Western Blot, Transfection, Knockdown

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: RIG-I-siRNA decreased HUA-induced fibrosis and pyroptosis molecules expressions in NRK-52E cells in vitro NRK-52E cells were transfected with RIG-I-siRNA or NC-siRNA and treated with UA for 48 h. (A–D) Western blot showing caspase-1 (A and B), GSDMD (A and C) and RIG-I (A and D) expressions after UA treatment with or without RIG-I knockdown (n = 6 per group). (E–G) Western blot showing cleaved-caspase-1 (E and F) and GSDMD-N (E and G) expressions after UA treatment with or without RIG-I knockdown (n = 6 per group). (H) IL-1β level in culture medium (n = 6 per group). (I–K) Western blot showing fibronectin (I and J) and α-SMA (I and K) expressions after UA treatment with or without RIG-I knockdown (n = 6 per group). Data for (B–D), (F–H) and (J–K) are presented as mean ± SEM. One-way ANOVA with Student-Newman-Keuls test for (B–D), (F–H) and (J–K). UA, uric acid.

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: In Vitro, Transfection, Western Blot, Knockdown

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet:

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: Recombinant, H&E Stain, Staining, Activity Assay, Transfection, Magnetic Beads, Enzyme-linked Immunosorbent Assay, Lysis, Bradford Protein Assay, Bicinchoninic Acid Protein Assay, Negative Control, Software, Fluorescence, Microscopy