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

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A-485 alleviates fibrosis and apoptosis in kidney by disrupting tandem activation of acetylation and phosphorylation on STAT3.

doi: 10.1016/j.biopha.2025.118217

Figure Lengend Snippet: Fig. 2. Effects of A-485 on the profibrotic phenotype induced by TGF-β1 in HK-2 cells. (A) The representative western blot images and quantification of FN, Col-I, E- cad and α-SMA in cultured HK-2 cells (n = 3). (B) Micrographs of α-SMA and E-cad immunofluorescence staining in HK-2 cells (n = 6). (C-D) The quantification of α-SMA and E-cad immunofluorescence staining in HK-2 cells. Values are expressed as means ± SEM. *p < 0.05, * *p < 0.01. FN, fibronectin; Col-I, collagen I; E-cad, E-Cadherin; α-SMA, α-smooth muscle actin.

Article Snippet: The antibodies used in the present study are listed as below: antibodies against Bcl-2 (ab196495), Collagen I (ab34710); antibodies against STAT3 (10253–2-AP), Bax (50599–2-Ig), α-SMA (14395–1-AP), Fibronectin (15613–1-AP), E-Cadherin (20874–1-AP) and GAPDH (10494–1-AP) (Proteintech, Chicago, IL, USA); antibodies against Acetyl-STAT3 (Lys685) (#2523), Cleaved Caspase-3 (#9661), p300 (#67621), CBP(#74384SF), Acetylated-Lysine (#9441), PhosphoSTAT3 (S727) (#49081) and Phospho-STAT3 (Y705) (#9145) (Cell Signaling Technology, Beverly, MA, USA); antibodies against NDUFS1 (ER1803–08) and SDHB (ER1803–63) (Hua’an Biotechnology, Hangzhou, China); antibody against COX4 (sc-517553) (Santa Cruz Biotechnology, Santa Cruz, CA); goat anti-rabbit or mouse IgG-HRP secondary antibodies, TRITC-labelled goat anti-rabbit secondary antibody and FITC-labelled goat anti-rabbit or mouse IgG secondary antibodies (Zhongshan Goldenbridge Biotechnology, Beijing, China); Inhibitor A-485 (1889279–16–6) (GlpBio, Montclair, USA).

Techniques: Western Blot, Cell Culture, Immunofluorescence, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A-485 alleviates fibrosis and apoptosis in kidney by disrupting tandem activation of acetylation and phosphorylation on STAT3.

doi: 10.1016/j.biopha.2025.118217

Figure Lengend Snippet: Fig. 4. Effects of A-485 on UUO-induced profibrotic proteins in mouse kidney. (A-B) The representative western blot images and quantification of FN, Col-I, E-cad and α-SMA in mice (n = 6). (C) Representative micrographs of FN, Col-I and E-cad immunohistochemical staining in mice. (D-F) The quantification of FN, Col-I and E- cad expression detected by immunohistochemical staining (n = 5). Values are expressed as means ± SEM. * *p < 0.01. sham, sham operation; UUO: unilateral ureteral obstruction; FN, fibronectin; Col-I, collagen I; E-cad, E-Cadherin; α-SMA, α-smooth muscle actin.

Article Snippet: The antibodies used in the present study are listed as below: antibodies against Bcl-2 (ab196495), Collagen I (ab34710); antibodies against STAT3 (10253–2-AP), Bax (50599–2-Ig), α-SMA (14395–1-AP), Fibronectin (15613–1-AP), E-Cadherin (20874–1-AP) and GAPDH (10494–1-AP) (Proteintech, Chicago, IL, USA); antibodies against Acetyl-STAT3 (Lys685) (#2523), Cleaved Caspase-3 (#9661), p300 (#67621), CBP(#74384SF), Acetylated-Lysine (#9441), PhosphoSTAT3 (S727) (#49081) and Phospho-STAT3 (Y705) (#9145) (Cell Signaling Technology, Beverly, MA, USA); antibodies against NDUFS1 (ER1803–08) and SDHB (ER1803–63) (Hua’an Biotechnology, Hangzhou, China); antibody against COX4 (sc-517553) (Santa Cruz Biotechnology, Santa Cruz, CA); goat anti-rabbit or mouse IgG-HRP secondary antibodies, TRITC-labelled goat anti-rabbit secondary antibody and FITC-labelled goat anti-rabbit or mouse IgG secondary antibodies (Zhongshan Goldenbridge Biotechnology, Beijing, China); Inhibitor A-485 (1889279–16–6) (GlpBio, Montclair, USA).

Techniques: Western Blot, Immunohistochemical staining, Staining, Expressing

Journal: eLife

Article Title: Tumor stiffening reversion through collagen crosslinking inhibition improves T cell migration and anti-PD-1 treatment

doi: 10.7554/elife.58688

Figure Lengend Snippet: Figure 5. Impact of LOX inhibition on T cell migration in EGI-1, MMTV-PyMT, mPDAC, and KPC tumor models. Migration of activated PBT plated onto fresh tumor slices was analyzed in EGI-1 and MMTV-PyMT tumor model, whilst resident tumor-infiltrating T lymphocytes were analyzed in mPDAC and KPC tumor model. Illustrative images of T cell migration tracks in EGI-1, MMTV-PyMT, mPDAC, and KPC tumor models. Tumor stroma (fibronectin) in red, tumor cells (EpCAM in EGI-1, MMTV-PyMT, and KPC tumor models, CD44 in mPDAC tumor models), in blue and T cells (CD8 in mPDAC and KPC, Calcein in MMTV-PyMT, and EGI-1 tumor models) in green. Tracks are color-coded to illustrate track displacement. Scale bar = 100 mm. T cell migration speed, T cell displacement, and trajectory straightness in all tumor models. ***p-value>0.001, p-value>0.05, Student’s t-test. Results are shown as mean ± SD. The online version of this article includes the following source data and figure supplement(s) for figure 5:

Article Snippet: The slices were then washed to remove all cells that had not infiltrated the slice and stained for 15 min at 37 ̊C with the following antibodies: BV421–anti- human EpCAM (9C4 clone; BioLegend) and anti-human/mouse fibronectin at 10 mg/mL (HFN7.1 clone; Acris antibodies).

Techniques: Inhibition, Migration

Journal: Renal Failure

Article Title: apoE Expression in Glomerulus and Correlation with Glomerulosclerosis Induced by Adriamycin in Rats

doi: 10.3109/0886022x.2011.559680

Figure Lengend Snippet: Figure 1. Glomerular morphology was normal in sham group (A1: 9 weeks; A2: 13 weeks; HE). Proliferation occurring in the majority of glomerular mesangial cells and extracellular matrix in GS group, and degeneration of glomerular epithelial cells and infiltration of wide- spread mononuclear cells were shown (A3: 9 weeks; A4: 13 weeks; HE), especially in 13 weeks of GS group (A4). Representative samples of immunohistochemical staining for glomerular Col-IV (SHO: B1: 9 weeks, B2: 13 weeks; GS: B3: 9 weeks and B4: 13 weeks), FN (SHO: C1: 9 weeks, C2: 13 weeks; GS: C3: 9 weeks and C4: 13 weeks), a-SMA (SHO: D1: 9 weeks, D2: 13 weeks; GS: D3: 9 weeks and D4: 13 weeks), TGF-b1 (SHO: E1: 9 weeks, E2: 13 weeks; GS: E3: 9 weeks and E4: 13 weeks), and apoE (SHO: F1: 9 weeks, F2: 13 weeks; GS: F3: 9 weeks and F4: 13 weeks) were observed in all groups. Sham group (B1, B2, C1, C2, D1, D2, E1, E2, F1, and F2): positive staining (in brown) was faint in glomerulus, glomerular endothelial cells, glomerular basement membrane, mesangial cells, and visceral epithelial cells. GS group (B3, B4, C3, C4, D3, D4, E3, E4, F3, and F4): positive staining was strong in most glomerulus, glomerular endothelial cells, glomerular basement membrane, mesangial cells, and visceral epithelial cells, especially in those of 13 weeks (B4, C4, D4, E4, and F4). Magnification 400×. Notes: SHO, sham operation group; GS, glomerulosclerosis model group; HE, hematoxylin and eosin; FN, fibronectin; a-SMA, a-smooth muscle actin; TGF-b1, transforming growth factor-b1; apoE, apolipoprotein E.

Article Snippet: Finally, the sections were incubated with primary monoclonal antibodies against apoE (1/150) (Bo Ao-Sen, Co., Beijing, China), Collagen IV (Col-IV; 1/100) (Shanghai ChangDao Co., Shanghai, China), fibronectin (FN; 1/ 100) (Beijing Zhongshan Co. Ltd., Beijing, China), a-smooth muscle actin (a-SMA) (ready-to-use kit) (Shanghai Changdao, Co.), and transforming growth factor-b1 (TGF-b1; 1/100) (Wuhan Boshide, Co., Wuhan, China) overnight at 4°C.

Techniques: Immunohistochemical staining, Staining, Membrane

Journal: Scientific reports

Article Title: C1QA and COMP: plasma-based biomarkers for early diagnosis of pancreatic neuroendocrine tumors.

doi: 10.1038/s41598-023-48323-x

Figure Lengend Snippet: Figure 5. Validation of potential protein targets. Western blot profiling and densitometric analysis of final selected proteins (A). Densitometric analysis of (B) C1QA (p = 0.001, 0.03, 0.0013); (C) COMP protein (p = 0.011, 0.019), (D) HSP90B1, (E) ITGA2B/CD41, (F) FN1, (G, H) Sandwich ELISA for C1QA binding analysis (I) ROC curve analysis for plasma C1QA levels in Grade I PanNET vs controls (J, K) Sandwich ELISA for COMP analysis (I) ROC curve analysis for plasma COMP levels in Grade I PanNET vs controls.

Article Snippet: Antibody-based validation of the C1QA (Affinity biosciences; DF7839), COMP (Affinity biosciences; DF13438), HSP90 (Affinity biosciences; AF0735), FN1 (Elabscience; E-AB-22077), and ITGA2B (Affinity biosciences; DF7456) was achieved by western blotting against their respective antibodies. β-actin (Abcam; ab8227) served as the internal control.

Techniques: Biomarker Discovery, Western Blot, Sandwich ELISA, Binding Assay, Clinical Proteomics