gs 1 inhibitor  (MedChemExpress)

Journal: Autoimmunity

Article Title: Increasing expression of dual-specificity phosphatase 12 mitigates oxygen-glucose deprivation/reoxygenation-induced neuronal apoptosis and inflammation through inactivation of the ASK1-JNK/p38 MAPK pathway.

doi: 10.1080/08916934.2024.2345919

Figure Lengend Snippet: Figure 6. The effect of DUSP12 overexpression on ASK1-JNK/p38 MAPK under OGD/R conditions. (A-D) The levels of phosphorylated ASK1, JNK and p38 in Ad-Ctrl- or Ad-DUSP12-infected neurons with or without OGD/R were determined by Western blotting and their protein quantification was shown. n = 3. **p < 0.01. Statistical differences were determined using one-way ANOVA followed by Tukey’s post-hoc test.

Article Snippet: Chemical inhibitor treatment The highly selective and potent ASK1 inhibitor GS-4997 (Selleck Chemicals, Shanghai, China) was utilized in this work.

Techniques: Over Expression, Infection, Western Blot

Journal: Autoimmunity

Article Title: Increasing expression of dual-specificity phosphatase 12 mitigates oxygen-glucose deprivation/reoxygenation-induced neuronal apoptosis and inflammation through inactivation of the ASK1-JNK/p38 MAPK pathway.

doi: 10.1080/08916934.2024.2345919

Figure Lengend Snippet: Figure 7. The effect of DUSP12 deficiency on ASK1-JNK/p38 MAPK under OGD/R conditions. (A-D) The levels of phosphorylated ASK1, JNK and p38 in Ad-sh- Scrambled- or Ad-sh-DUSP12-infected neurons with or without OGD/R were determined by Western blotting and their protein quantification was shown. n = 3. **p < 0.01. Statistical differences were determined using one-way ANOVA followed by Tukey’s post-hoc test.

Article Snippet: Chemical inhibitor treatment The highly selective and potent ASK1 inhibitor GS-4997 (Selleck Chemicals, Shanghai, China) was utilized in this work.

Techniques: Infection, Western Blot

Journal: Autoimmunity

Article Title: Increasing expression of dual-specificity phosphatase 12 mitigates oxygen-glucose deprivation/reoxygenation-induced neuronal apoptosis and inflammation through inactivation of the ASK1-JNK/p38 MAPK pathway.

doi: 10.1080/08916934.2024.2345919

Figure Lengend Snippet: Figure 8. The effect of ASK1 inhibition on DUSP12-mediated JNK/p38 MAPK. Ad-sh-DUSP12-infected neurons were treated with the ASK1 inhibitor GS-4997 and subjected to OGD/R. (A, D) Levels of DUSP12 and phosphorylated ASK1, JNK and p38 were examined by Western blotting and their protein quantification was shown. n = 3. **p < 0.01 and #p > 0.05. Statistical differences were determined using one-way ANOVA followed by Tukey’s post-hoc test.

Article Snippet: Chemical inhibitor treatment The highly selective and potent ASK1 inhibitor GS-4997 (Selleck Chemicals, Shanghai, China) was utilized in this work.

Techniques: Inhibition, Infection, Western Blot

Journal: Autoimmunity

Article Title: Increasing expression of dual-specificity phosphatase 12 mitigates oxygen-glucose deprivation/reoxygenation-induced neuronal apoptosis and inflammation through inactivation of the ASK1-JNK/p38 MAPK pathway.

doi: 10.1080/08916934.2024.2345919

Figure Lengend Snippet: Figure 9. The effect of ASK1 inhibition on DUSP12-mediated OGD/R injury. Neuronal apoptosis in different groups was monitored by the (A, B) TUNEL (scale bar = 100 μm) and (C, D) Annexin V-FITC/PI staining assays. (E, F) Concentrations of TNF-α, IL-6 and IL-1β in different groups were quantified by ELISA. n = 3. **p < 0.01 and #p > 0.05. Statistical differences were determined using one-way ANOVA followed by Tukey’s post-hoc test.

Article Snippet: Chemical inhibitor treatment The highly selective and potent ASK1 inhibitor GS-4997 (Selleck Chemicals, Shanghai, China) was utilized in this work.

Techniques: Inhibition, TUNEL Assay, Staining, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Biological Sciences

Article Title: Dual-specificity Phosphatase 9 protects against Cardiac Hypertrophy by targeting ASK1

doi: 10.7150/ijbs.57130

Figure Lengend Snippet: DUSP9 modulated cardiac hypertrophy via the ASK1-p38/JNK1/2 signaling axis. (A, B) Representative western blots and quantification of both phosphorylated and total protein levels of ASK1, p38, ERK1/2 and JNK in the hearts of control and DUSP9-CKO (A) or NTG and DUSP9-TG mice (B) four weeks after TAC treatment or sham (n = 4 mice per group) (C, D) Representative western blots and quantification of phosphorylated and total protein levels of ASK1, p38, ERK and JNK in NRVM with DUSP9 knockdown (C) or DUSP9 overexpression ( D ) after 48 h of PBS or Ang II (1 µM) challenge. Data are presented as the mean ±SD. *P<0.05, **P<0.01, n.s. means no significant difference; NRVM refers to neonatal rat ventricular myocytes.

Article Snippet: The cells undergone starvation under serum-free medium condition for 12 h to synchronize before treatment with Ang II (1 μM) or PBS or ASK1 inhibitor GS-4997(iASK1 80 μM, 1148428-04-3, Selleck) or DMSO for another 48 h.

Techniques: Western Blot, Control, Knockdown, Over Expression

Journal: International Journal of Biological Sciences

Article Title: Dual-specificity Phosphatase 9 protects against Cardiac Hypertrophy by targeting ASK1

doi: 10.7150/ijbs.57130

Figure Lengend Snippet: DUSP9 regulated Ang II-induced cardiac hypertrophy by binding to ASK1. (A) Immunofluorescence staining of DUSP9 (red), ASK1 (green) and nuclei (blue) in HEK293T cells. (B) Co-immunoprecipitation assay was performed in HEK293T cells that co-transfected with Flag-DUSP9 and HA-ASK1 to measure the interaction between DUSP9 and ASK1. (C) Co-immunoprecipitation assay in NRVM infected with Flag-DUSP9 to exam the interaction between DUSP9 and endogenous ASK1. (D) Western blots of DUSP9 and phosphorylated ASK1 protein level in DUSP9 knockdown NRVM treated with DMSO or iASK1 along with Ang II (1 µM) insult for 48 h. (E, F) Representative fluorescent images and quantification of cell surface area of DUSP9 knockdown NRVM with DMSO or iASK1 with Ang II (1 µM) stimulation for 48 h (Scale bar = 20 µm) (G) mRNA levels of hypertrophic biomarkers ( Anp and Myh7 ) in DUSP9 knockdown NRVM undergoing Ang II (1 µM) treatment for 48 h along with DMSO or iASK1. Data are presented as the mean ±SD. *P<0.05, **P<0.01; NRVM refers to neonatal rat ventricular myocytes.

Article Snippet: The cells undergone starvation under serum-free medium condition for 12 h to synchronize before treatment with Ang II (1 μM) or PBS or ASK1 inhibitor GS-4997(iASK1 80 μM, 1148428-04-3, Selleck) or DMSO for another 48 h.

Techniques: Binding Assay, Immunofluorescence, Staining, Co-Immunoprecipitation Assay, Transfection, Infection, Western Blot, Knockdown

Journal: Pulmonary Circulation

Article Title: Apoptosis signal-regulating kinase 1 inhibition in in vivo and in vitro models of pulmonary hypertension

doi: 10.1177/2045894020922810

Figure Lengend Snippet: Sugen/hypoxic rat cardiopulmonary cardiac characteristics after ASK1 inhibition. The impact of ASK1 inhibitor GS-44217 in reversing established pulmonary hypertension (PH) was investigated in the Sugen/hypoxic (SuHx) rat model. Four groups of animals were investigated; SuHx animals treated with vehicle (SuHx + V) or ASK1 inhibitor GS-444217 (SuHx + GS) for three weeks post disease establishment (at five weeks) or age-matched normoxic controls treated with vehicle (Norm + V) or drug (Norm + GS). (a) Pulmonary vascular remodelling was investigated through analysis of high-power magnification images of lung alpha smooth muscle actin (α-SMA) immunohistochemistry, arrows show pulmonary vessels of interest and (b) and pulmonary vascular remodelling quantification. (c–f) Protein abundance determined and quantified by Western blot analysis in tissue homogenate for phospho p38 MAPK (pp38 MAPK) in (c) lung, (d) right ventricle (RV), (e) left ventricle (LV) and (f) phospho ASK1 (pASK1) in lung. All protein abundance is normalised to total p38 MAPK (tp38 MAPK) and is presented as relative abundance (AU). Data are n = 4 biological replicates per group and are displayed as mean ± SEM analysed by one-way ANOVA with Turkey's post hoc analysis. * p ≤ 0.05. Plasma analysis was also carried out to quantify the levels of (g) endothelin-1 (ET-1), (h) tissue inhibitor of metalloproteinases (TIMP1) and (i) B-type natriuretic peptide (BNP). Quantification data is n = 5 per group for all plasma analyses and displayed as individual data points with mean. Statistical analysis is by one-way ANOVA with Turkey's post hoc analysis * p ≤ 0.05 and ** p ≤ 0.01. MAPK: mitogen-activated protein kinase; ASK1: apoptosis signal-regulating kinase 1.

Article Snippet: This study was funded as part of an industrial collaboration with Gilead Science Inc. Apoptosis signal-regulating kinase 1 inhibitor GS-444217 was provided by Gilead Science Inc.

Techniques: Inhibition, Immunohistochemistry, Quantitative Proteomics, Western Blot, Clinical Proteomics

Journal: Pulmonary Circulation

Article Title: Apoptosis signal-regulating kinase 1 inhibition in in vivo and in vitro models of pulmonary hypertension

doi: 10.1177/2045894020922810

Figure Lengend Snippet: ASK1 inhibition prevents hypoxia-induced increases in protein abundance in rat pulmonary artery fibroblasts (RPAFs) derived from wild-type animals. RPAF were exposed to either normoxic or hypoxic culture conditions for 24 h in the presence or absence of 1% serum (+serum), with or without the ASK1 inhibitor GS-444217 (+GS), protein abundance was determined in the resulting cell lysate by Western blot analysis. (a) Typical Western blots of the hypoxia-induced transcription factor HIF-1α, phosphorylated (p) ASK1, MKK3/6, MKK4, JNK and p38 MAPK. Total (t) p38 MAPK is used here as a house-keeping protein and loading control for densitometry of Western blots to determine protein relative abundance. Protein abundance was graphically represented as bar chart (b) HIF-1α, (c) pASK1, (d) pERK1/2, (e) pp 38 MAPK, (f) pMKK3/6 and (g) pJNK. n = 3 (biological replicates), displayed as bar chart as mean ± SEM for ease of interpretation. All stats are vs normoxic controls analysed by one-way ANOVA and Bonferroni's multiple comparison test. * p ≤ 0.05 and ** p ≤ 0.01. Cell culture conditioned media analysis was also carried out to quantify the levels of (h) soluble intracellular adhesion molecule 1 (sICAM-1), (i) tissue inhibitor of metalloproteinases (TIMP1) and (j) endothelin-1 (ET-1). Quantification data are n = 5 per group for all plasma analyses and displayed as individual data points with error bars representing mean ± SEM. Statistical analysis is by one-way ANOVA with Turkey's post hoc analysis. * p ≤ 0.05 and ** p ≤ 0.01. SFM: serum free media; HIF-1α: hypoxia-inducible factor-1α; pASK1: phospho apoptosis signal-regulating kinase 1; pp 38 MAPK: phospho p38 mitogen-activated protein kinase.

Article Snippet: This study was funded as part of an industrial collaboration with Gilead Science Inc. Apoptosis signal-regulating kinase 1 inhibitor GS-444217 was provided by Gilead Science Inc.

Techniques: Inhibition, Quantitative Proteomics, Derivative Assay, Western Blot, Control, Comparison, Cell Culture, Clinical Proteomics