stat5 sirna  (Santa Cruz Biotechnology)

Journal: Heliyon

Article Title: Insulin augments angiotensin II-induced myocardial fibrosis via the MEK/STAT3 pathway

doi: 10.1016/j.heliyon.2023.e22860

Figure Lengend Snippet: Insulin facilitated Ang II-mediated STAT expression and activation in rat heart tissue. Protein levels of p-STAT1, STAT1, p-STAT3, STAT3, p-STAT5 and STAT5 in the heart tissues of insulin-treated rats with or without Ang II-induced heart failure were detected by WB analysis, and the phosphorylation of STAT1, 3, and 5 was determined by calculating the ratio of p-STATs/STATs. The original image of Western blot is in the supplementary material. *, P < 0.05 compared with the control group; Data are expressed means ± SEM, determined in three independent experiments. Oneway ANOVA, followed by LSD t and SNK post hoc test, was performed to analyze difference between the groups. ***, P < 0.001. #, P < 0.05 compared to the insulin group, ##, P < 0.01; ###, P < 0.001. &, P < 0.05 compared with the Ang II group, &&, P < 0.01; &&&, P < 0.001.

Article Snippet: STAT3 and STAT5 silencing was performed by transfection with small interfering RNAs (siRNAs) against STAT3 and STAT5 (sc-29494 and sc-29496; Santa Cruz Biotechnology, USA) as described in our previous study [ ].

Techniques: Expressing, Activation Assay, Phospho-proteomics, Western Blot, Control

Journal: Heliyon

Article Title: Insulin augments angiotensin II-induced myocardial fibrosis via the MEK/STAT3 pathway

doi: 10.1016/j.heliyon.2023.e22860

Figure Lengend Snippet: Insulin-mediated promotion of AngII-induced proliferation and collagen synthesis in rat CFs is dependent on the upregulation of STAT3. A, B: Primary rat cardiac fibroblasts were transfected with invalid, STAT5, or STAT3 siRNA for 24 h and were then exposed to 100 nmol/L Ang II and/or 100 nmol/L insulin for another 24 h. The protein levels of p-STAT3, STAT3 (A), p-STAT5 and STAT5 (B) were measured by WB analysis. C: A [3H]-thymidine incorporation assay was used to measure the proliferation of CFs. D: qPCR analysis of the mRNA expression of the fibrotic gene collagen COL1A1. The original image of Western blot is in the supplementary material. Data are expressed means ± SEM, determined in three independent experiments. Oneway ANOVA, followed by LSD t and SNK post hoc test, was performed to analyze difference between the groups. ***, P < 0.001, compared with the control group; ###compared with the Ang II group, P < 0.001.

Article Snippet: STAT3 and STAT5 silencing was performed by transfection with small interfering RNAs (siRNAs) against STAT3 and STAT5 (sc-29494 and sc-29496; Santa Cruz Biotechnology, USA) as described in our previous study [ ].

Techniques: Transfection, Thymidine Incorporation Assay, Expressing, Western Blot, Control

Journal: Heliyon

Article Title: Insulin augments angiotensin II-induced myocardial fibrosis via the MEK/STAT3 pathway

doi: 10.1016/j.heliyon.2023.e22860

Figure Lengend Snippet: Insulin facilitated Ang II-mediated STAT expression and activation in rat heart tissue. Protein levels of p-STAT1, STAT1, p-STAT3, STAT3, p-STAT5 and STAT5 in the heart tissues of insulin-treated rats with or without Ang II-induced heart failure were detected by WB analysis, and the phosphorylation of STAT1, 3, and 5 was determined by calculating the ratio of p-STATs/STATs. The original image of Western blot is in the supplementary material. *, P < 0.05 compared with the control group; Data are expressed means ± SEM, determined in three independent experiments. Oneway ANOVA, followed by LSD t and SNK post hoc test, was performed to analyze difference between the groups. ***, P < 0.001. #, P < 0.05 compared to the insulin group, ##, P < 0.01; ###, P < 0.001. &, P < 0.05 compared with the Ang II group, &&, P < 0.01; &&&, P < 0.001.

Article Snippet: STAT3 and STAT5 silencing was performed by transfection with small interfering RNAs (siRNAs) against STAT3 and STAT5 (sc-29494 and sc-29496; Santa Cruz Biotechnology, USA) as described in our previous study [ ].

Techniques: Expressing, Activation Assay, Phospho-proteomics, Western Blot, Control

Journal: Heliyon

Article Title: Insulin augments angiotensin II-induced myocardial fibrosis via the MEK/STAT3 pathway

doi: 10.1016/j.heliyon.2023.e22860

Figure Lengend Snippet: Insulin-mediated promotion of AngII-induced proliferation and collagen synthesis in rat CFs is dependent on the upregulation of STAT3. A, B: Primary rat cardiac fibroblasts were transfected with invalid, STAT5, or STAT3 siRNA for 24 h and were then exposed to 100 nmol/L Ang II and/or 100 nmol/L insulin for another 24 h. The protein levels of p-STAT3, STAT3 (A), p-STAT5 and STAT5 (B) were measured by WB analysis. C: A [3H]-thymidine incorporation assay was used to measure the proliferation of CFs. D: qPCR analysis of the mRNA expression of the fibrotic gene collagen COL1A1. The original image of Western blot is in the supplementary material. Data are expressed means ± SEM, determined in three independent experiments. Oneway ANOVA, followed by LSD t and SNK post hoc test, was performed to analyze difference between the groups. ***, P < 0.001, compared with the control group; ###compared with the Ang II group, P < 0.001.

Article Snippet: STAT3 and STAT5 silencing was performed by transfection with small interfering RNAs (siRNAs) against STAT3 and STAT5 (sc-29494 and sc-29496; Santa Cruz Biotechnology, USA) as described in our previous study [ ].

Techniques: Transfection, Thymidine Incorporation Assay, Expressing, Western Blot, Control

Journal: Heliyon

Article Title: Insulin augments angiotensin II-induced myocardial fibrosis via the MEK/STAT3 pathway

doi: 10.1016/j.heliyon.2023.e22860

Figure Lengend Snippet: The effect of insulin on STAT3 expression and AngII-induced proliferation and collagen synthesis in rat CFs was mediated by the MEK pathway. Primary rat cardiac fibroblasts were pretreated with the PI3K inhibitor LY294002 (10 μmol/L) or MEK inhibitor PD0325901 (1 μmol/L) for 30 min and then exposed to 100 nmol/L Ang II and/or 100 nmol/L insulin for 24 h. A, B. The protein levels of p-STAT3 and STAT3 were measured by WB analysis. C: A [3H]-thymidine incorporation assay was used to measure the proliferation of CFs. D: qPCR analysis of the mRNA expression of the fibrotic gene collagen COL1A1. The original image of Western blot is in the supplementary material. Data are expressed means ± SEM, determined in three independent experiments. Oneway ANOVA, followed by LSD t and SNK post hoc test, was performed to analyze difference between the groups. Significantly different from control, **, P < 0.01; ***, P < 0.001; compared with the Ang II (or Ang II + LY) group, #, P < 0.05; ##, P < 0.01; ###, P < 0.001.

Article Snippet: STAT3 and STAT5 silencing was performed by transfection with small interfering RNAs (siRNAs) against STAT3 and STAT5 (sc-29494 and sc-29496; Santa Cruz Biotechnology, USA) as described in our previous study [ ].

Techniques: Expressing, Thymidine Incorporation Assay, Western Blot, Control

Journal: Heliyon

Article Title: Insulin augments angiotensin II-induced myocardial fibrosis via the MEK/STAT3 pathway

doi: 10.1016/j.heliyon.2023.e22860

Figure Lengend Snippet: Insulin facilitated Ang II-mediated STAT expression and activation in rat heart tissue. Protein levels of p-STAT1, STAT1, p-STAT3, STAT3, p-STAT5 and STAT5 in the heart tissues of insulin-treated rats with or without Ang II-induced heart failure were detected by WB analysis, and the phosphorylation of STAT1, 3, and 5 was determined by calculating the ratio of p-STATs/STATs. The original image of Western blot is in the supplementary material. *, P < 0.05 compared with the control group; Data are expressed means ± SEM, determined in three independent experiments. Oneway ANOVA, followed by LSD t and SNK post hoc test, was performed to analyze difference between the groups. ***, P < 0.001. #, P < 0.05 compared to the insulin group, ##, P < 0.01; ###, P < 0.001. &, P < 0.05 compared with the Ang II group, &&, P < 0.01; &&&, P < 0.001.

Article Snippet: STAT3 and STAT5 silencing was performed by transfection with small interfering RNAs (siRNAs) against STAT3 and STAT5 (sc-29494 and sc-29496; Santa Cruz Biotechnology, USA) as described in our previous study [ ].

Techniques: Expressing, Activation Assay, Phospho-proteomics, Western Blot, Control

Journal: Heliyon

Article Title: Insulin augments angiotensin II-induced myocardial fibrosis via the MEK/STAT3 pathway

doi: 10.1016/j.heliyon.2023.e22860

Figure Lengend Snippet: Insulin-mediated promotion of AngII-induced proliferation and collagen synthesis in rat CFs is dependent on the upregulation of STAT3. A, B: Primary rat cardiac fibroblasts were transfected with invalid, STAT5, or STAT3 siRNA for 24 h and were then exposed to 100 nmol/L Ang II and/or 100 nmol/L insulin for another 24 h. The protein levels of p-STAT3, STAT3 (A), p-STAT5 and STAT5 (B) were measured by WB analysis. C: A [3H]-thymidine incorporation assay was used to measure the proliferation of CFs. D: qPCR analysis of the mRNA expression of the fibrotic gene collagen COL1A1. The original image of Western blot is in the supplementary material. Data are expressed means ± SEM, determined in three independent experiments. Oneway ANOVA, followed by LSD t and SNK post hoc test, was performed to analyze difference between the groups. ***, P < 0.001, compared with the control group; ###compared with the Ang II group, P < 0.001.

Article Snippet: STAT3 and STAT5 silencing was performed by transfection with small interfering RNAs (siRNAs) against STAT3 and STAT5 (sc-29494 and sc-29496; Santa Cruz Biotechnology, USA) as described in our previous study [ ].

Techniques: Transfection, Thymidine Incorporation Assay, Expressing, Western Blot, Control