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: Mouse Fibronectin primer , Sango biotech , Decribed in current manuscript.

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: Mouse Fibronectin primer , Sango biotech , Decribed in current manuscript.

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: Mouse Fibronectin primer , Sango biotech , Decribed in current manuscript.

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: Mouse Fibronectin primer , Sango biotech , Decribed in current manuscript.

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: Mouse Fibronectin primer , Sango biotech , Decribed in current manuscript.

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: iScience

Article Title: Ym1 + macrophages orchestrate fibrosis, lesion growth, and progression during development of murine pancreatic cancer

doi: 10.1016/j.isci.2022.104327

Figure Lengend Snippet: Pancreatic Ym1 + macrophages produce TGFβ1 (A and B) Macrophages were isolated from murine bone marrow (BMDM) or peritoneal cavity (PM) and polarized using 10 ng/mL LPS and 20 ng/mL IFNγ (M1) or 20 ng/mL IL-4 (M2). qPCR analysis indicates Ym1 + M2 macrophages produce Tgfb1 /TGFβ1. Error bars represent the standard deviation and using the t-test, p-values indicate statistical significance. (C) IF-IHC for Ym1 and TGFβ1(Y369) with nuclei stained by DAPI. Subsequently, an H&E staining was done. Scale bar is 50 μm. (D and E) Pancreas tissue samples were examined for expression of TGFβ1(Y369). Representative images are shown with 50 μm scale bars (D). Quantification of TGFβ1(Y369) was done on whole tissue slide area of n = 5 control or n = 3 Ym1 + macrophage-depleted tissues. Error bars represent the standard deviation and using the t-test, p-values indicate statistical significance.

Article Snippet: qPCR primer/probe: Fn1 /Fibronectin , Thermo Fisher Scientific , Mm01256744_m1.

Techniques: Isolation, Standard Deviation, Staining, Expressing, Control

Journal: iScience

Article Title: Ym1 + macrophages orchestrate fibrosis, lesion growth, and progression during development of murine pancreatic cancer

doi: 10.1016/j.isci.2022.104327

Figure Lengend Snippet: TGFβ1 activates quiescent PSCs and increases SMA and TIMP1 expression in activated, αSMA-positive, fibroblast-like PSCs (A) Brightfield images of freshly isolated qPSCs treated with TGFβ1 or vehicle control for 48 h. Scale bar indicates 100 μm. (B) qPCR analysis of aflPSC markers in freshly isolated PSCs treated with TGFβ1 or vehicle control for 48 h. Error bars represent the standard deviation. The t-test indicates statistical significance. (C): Acta2 /αSMA mRNA expression in response to treatment of aflPSCs with TGFβ1. Error bars represent the standard deviation, and the t-test indicates statistical significance. (D) Array to detect cytokines secreted by aflPSCs. (E) Proliferation assay for aflPSCs treated with CXCL12 neutralizing antibody (NAB) or isotype control. Error bars represent the standard deviation, and the t-test indicates statistical significance.

Article Snippet: qPCR primer/probe: Fn1 /Fibronectin , Thermo Fisher Scientific , Mm01256744_m1.

Techniques: Expressing, Isolation, Control, Standard Deviation, Proliferation Assay

Journal: iScience

Article Title: Ym1 + macrophages orchestrate fibrosis, lesion growth, and progression during development of murine pancreatic cancer

doi: 10.1016/j.isci.2022.104327

Figure Lengend Snippet: TIMP1 expression is enhanced by TGFβ1 signaling in aflPSCs (A) Brightfield images of PSCs treated with recombinant TIMP1 or vehicle control. The scale bar indicates 100 μm. (B) qPCR analysis for Timp1 in PanIN cells, aflPSCs, Ym1 + BMDM, and Ym1 + PM. Statistical significance determined by the t-test and error bars represent the standard deviation. (C) In situ hybridization (ISH) for Timp1 mRNA in brown combined with IHC for Ym1 protein in pink on KC mouse tissue. Scale bar indicates 100 μm. (D) qPCR analysis of Timp1 in aflPSCs treated with TGFβ1 or vehicle. Error bars represent standard deviation and the t-test was used to determine statistical significance. (E) IHC for TIMP1 protein (brown) in control or Ym1 + macrophage-depleted KC tissue. Scale bars indicate 50 μm. (F) Pancreas tissue from p48 cre ;LSL-Kras G12D mice treated with mIL-13 neutralizing antibody or isotype matching control antibody was stained for SMA (red) and TIMP1 (green). Nuclei were visualized using DAPI and scale bars indicate 50 μm.

Article Snippet: qPCR primer/probe: Fn1 /Fibronectin , Thermo Fisher Scientific , Mm01256744_m1.

Techniques: Expressing, Recombinant, Control, Standard Deviation, In Situ Hybridization, Staining

Journal: iScience

Article Title: Ym1 + macrophages orchestrate fibrosis, lesion growth, and progression during development of murine pancreatic cancer

doi: 10.1016/j.isci.2022.104327

Figure Lengend Snippet: TIMP1 increases growth of LG-PanINs via its receptor CD63 (A) Representative picture of an IHC for CD63 (brown) in normal acinar, stromal, and lesion areas of KC tissue. Scale bars indicate 50 μm. (B) Quantification of CD63 expression in acinar, stromal, and lesion areas (n = 15 each). Points are graphed in a box and whiskers plot, where the whiskers extend to the minimum and maximum and the midline of the box indicates the median. Statistical significance was determined by a one-way ANOVA(p < 0.0001) followed by t-tests, p-values noted in figure. (C) qPCR for TGFβ1 or vehicle-treated PanIN organoids. Error bars represent the standard deviation and statistical significance was determined by the t-test. (D) TIMP1 increases the growth of duct-like structures. PanIN cells were seeded on top of Matrigel and treated with vehicle control or TIMP1 (50 ng/mL) for 4 days. Pictures show a representative area of single cells seeded at day 0 or ductal structures developed at day 4. The scale bar indicates 50 μm. (E) Quantification of areas of ductal structures in control (n = 68) or TIMP1 (n = 63)-stimulated organoids on day 4. For statistical analysis of data, a t-test was performed. The p-value (p = 0.0043) indicates statistical significance. The line indicates the median. (F) Quantification of areas of ductal structures in TIMP1-stimulated organoids that were incubated with 1 μg/mL control (IgG) or CD63 neutralization antibody (NOVUS, NBP2-42225SS). On day 4, size of ducts was analyzed (n = 276 for control; n = 404 for CD63-NAB). For statistical analysis of data, a t-test was performed. The p-value (p < 0.0001) indicates statistical significance. The line indicates the median.

Article Snippet: qPCR primer/probe: Fn1 /Fibronectin , Thermo Fisher Scientific , Mm01256744_m1.

Techniques: Expressing, Standard Deviation, Control, Incubation, Neutralization

Journal: iScience

Article Title: Ym1 + macrophages orchestrate fibrosis, lesion growth, and progression during development of murine pancreatic cancer

doi: 10.1016/j.isci.2022.104327

Figure Lengend Snippet: TGFβ1 drives EMT-like structural changes in PanIN Cells (A) PanIN organoids were treated with vehicle control or TGFβ1 three days after plating on top of Matrigel, and then imaged on days one and five post treatment. Scale bars indicate 100 μm. (B) qPCR analysis for Cdh1 (e-cadherin) and Cdh2 (n-cadherin) in vehicle or TGFβ1-treated PanIN organoids. Error bars indicate standard deviation and the t-test was used to determine statistical significance. (C) Left side: IHC for SMAD4 (brown) in KC mouse tissue. Images show collapsed PanIN structures (top, arrows) and non-collapsed PanIN structures (bottom image). Scale bars indicate 100 μm. Right side: Quantification of SMAD4 expression in non-collapsed (n = 46) and collapsed PanIN (n = 45) areas. Points are graphed in a box and whiskers plot, where the whiskers extend to the minimum and maximum and the midline of the box indicates the median. Statistical significance was determined by a t-test. (D) Tissue from KC mice was stained for N-cadherin (pink) and SMAD4 (green). DAPI was used to visualize nuclei. The scale bars indicate 50 μm.

Article Snippet: qPCR primer/probe: Fn1 /Fibronectin , Thermo Fisher Scientific , Mm01256744_m1.

Techniques: Control, Standard Deviation, Expressing, Staining

Journal: iScience

Article Title: Ym1 + macrophages orchestrate fibrosis, lesion growth, and progression during development of murine pancreatic cancer

doi: 10.1016/j.isci.2022.104327

Figure Lengend Snippet:

Article Snippet: qPCR primer/probe: Fn1 /Fibronectin , Thermo Fisher Scientific , Mm01256744_m1.

Techniques: Control, Recombinant, CyQUANT Assay, Proliferation Assay, RNAscope, Software, Microscopy, Real-time Polymerase Chain Reaction, In Situ Hybridization

Journal: Antioxidants

Article Title: High-Intensity Interval Training and Moderate-Intensity Continuous Training Attenuate Oxidative Damage and Promote Myokine Response in the Skeletal Muscle of ApoE KO Mice on High-Fat Diet

doi: 10.3390/antiox10070992

Figure Lengend Snippet: Description of primers used for quantitative real-time PCR.

Article Snippet: In addition, the primer sequences of glutamate-cysteine ligase modifier subunit ( Gclm ), glutathione synthase ( Gss ), NADPH oxidase 2 ( Nox2 ) (also called gp91phox ), neutrophil cytosolic factor 1 ( Ncf1 ) (also called p47phox ), NADPH oxidase 4 ( Nox4 ), fibronectin type III domain containing 5 ( Fndc5 ), acyl-CoA dehydrogenase short chain ( Acads ), hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha ( Hadha ), and hydroxyacyl-coenzyme A dehydrogenase (Hadh) were listed in and these primers were synthesized by Invitrogen Trading Co., Ltd. (Shanghai, China).

Techniques:

Journal: Antioxidants

Article Title: High-Intensity Interval Training and Moderate-Intensity Continuous Training Attenuate Oxidative Damage and Promote Myokine Response in the Skeletal Muscle of ApoE KO Mice on High-Fat Diet

doi: 10.3390/antiox10070992

Figure Lengend Snippet: Effects of HIIT and MICT on the mRNA expression of Fndc5 ( A ), Hadh, Acads, and Hadha ( D – F ), and musclin level ( C ) in the gastrocnemius as well as plasma irisin level ( B ) of WT and ApoE KO mice with HFD. Values are displayed as the mean ± SEM (n = 10/group). # p < 0.05 vs. ApoE −/ − CON; ## p < 0.01 vs. ApoE −/ − CON; †† p < 0.01 vs. ApoE −/ − MICT.

Article Snippet: In addition, the primer sequences of glutamate-cysteine ligase modifier subunit ( Gclm ), glutathione synthase ( Gss ), NADPH oxidase 2 ( Nox2 ) (also called gp91phox ), neutrophil cytosolic factor 1 ( Ncf1 ) (also called p47phox ), NADPH oxidase 4 ( Nox4 ), fibronectin type III domain containing 5 ( Fndc5 ), acyl-CoA dehydrogenase short chain ( Acads ), hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha ( Hadha ), and hydroxyacyl-coenzyme A dehydrogenase (Hadh) were listed in and these primers were synthesized by Invitrogen Trading Co., Ltd. (Shanghai, China).

Techniques: Expressing