Journal: Frontiers in cardiovascular medicine

Article Title: Inhibition of Peptidyl Arginine Deiminase 4-Dependent Neutrophil Extracellular Trap Formation Reduces Angiotensin II-Induced Abdominal Aortic Aneurysm Rupture in Mice.

doi: 10.3389/fcvm.2021.676612

Figure Lengend Snippet: FIGURE 6 | NETs induced VSMC apoptosis via p38/JNK pathway. Primary VSMCs isolated from aortas of ApoE−/−mice were treated with vehicle or 10 µg NETs for 24 h. (A) TUNEL staining for apoptosis cells (red). The percentage of apoptosis cells was analyzed. VSMCs were stained with SM22α. Nuclei were stained with DAPI. n = 3/vehicle group, n = 4/NET group, **p < 0.01. Statistical significance was analyzed by the unpaired t-test. (B) Annexin V-FITC/PI staining for VSMC apoptosis. n = 4/vehicle group, n = 3/NET group, *p < 0.05. Statistical significance was analyzed by the unpaired t-test. (C) Western blot for p38/JNK level. n = 3–4, *p < 0.05, ***p < 0.001. Statistical significance was analyzed by the unpaired t-test.

Article Snippet: To determine whether p38 inhibitor SB203580 or JNK inhibitor SP600125 could reverse NET-induced VSMC apoptosis, p38 inhibitor SB203580 (S1076, Selleckchem, Houston, TX, USA) or JNK inhibitor SP600125 (S1876, Beyotime, Shanghai, China) was preincubated in primary VSMC.

Techniques: Isolation, TUNEL Assay, Staining, Western Blot

Journal: Frontiers in cardiovascular medicine

Article Title: Inhibition of Peptidyl Arginine Deiminase 4-Dependent Neutrophil Extracellular Trap Formation Reduces Angiotensin II-Induced Abdominal Aortic Aneurysm Rupture in Mice.

doi: 10.3389/fcvm.2021.676612

Figure Lengend Snippet: FIGURE 7 | Inhibition p38/JNK pathway suppressed NETs induced VSMC apoptosis. Primary VSMCs isolated from aortas of ApoE−/−mice were preincubated with p38 inhibitor SB203580 or JNK inhibitor SP600125 for 30 min before NET treatment for 24 h. (A) TUNEL staining for apoptosis cells (red) and the percentage of apoptosis cells were analyzed. VSMCs were stained with SM22α. Nuclei were stained with DAPI. n = 3 per group, *p < 0.05, **p < 0.01. Statistical significance was determined by one-way ANOVA test followed by the unpaired t-test.

Article Snippet: To determine whether p38 inhibitor SB203580 or JNK inhibitor SP600125 could reverse NET-induced VSMC apoptosis, p38 inhibitor SB203580 (S1076, Selleckchem, Houston, TX, USA) or JNK inhibitor SP600125 (S1876, Beyotime, Shanghai, China) was preincubated in primary VSMC.

Techniques: Inhibition, Isolation, TUNEL Assay, Staining

Journal: Diabetes

Article Title: Altered MAPK Signaling in Progressive Deterioration of Endothelial Function in Diabetic Mice

doi: 10.2337/db12-0559

Figure Lengend Snippet: Shear stress (20 dynes/cm 2 )–induced dilation in mesenteric arteries of 9M Lepr db−/− and Lepr db+/− mice, in control and in the presence of resveratrol. n = 6–8 per group. *Significant difference between groups. PD, passive diameter.

Article Snippet: In separate experiments, SSID was assessed in vessels of 9M mice in control and in the presence of resveratrol (100 nmol/L, Erk activator; 3,4',5-trihydroxystilbene; LKT Laboratories) and resveratrol plus one of following inhibitors: SB203580, SP600125, and VAS2870 without or with additional U0126 (1 μmol/L, inhibitor of Erk).

Techniques: Shear, Control

Journal: Diabetes

Article Title: Altered MAPK Signaling in Progressive Deterioration of Endothelial Function in Diabetic Mice

doi: 10.2337/db12-0559

Figure Lengend Snippet: Shear stress (20 dynes/cm 2 )–induced dilation in mesenteric arteries of 9M Lepr db−/− mice in control and in the presence (+) or absence (─) of resveratrol, SB203580, SP600125, VAS2870, and U0126, respectively. n = 8 per group. *Significant difference between groups. PD, passive diameter.

Article Snippet: In separate experiments, SSID was assessed in vessels of 9M mice in control and in the presence of resveratrol (100 nmol/L, Erk activator; 3,4',5-trihydroxystilbene; LKT Laboratories) and resveratrol plus one of following inhibitors: SB203580, SP600125, and VAS2870 without or with additional U0126 (1 μmol/L, inhibitor of Erk).

Techniques: Shear, Control

Journal: Scientific Reports

Article Title: Upregulated long noncoding RNA LOC105375913 induces tubulointerstitial fibrosis in focal segmental glomerulosclerosis

doi: 10.1038/s41598-018-36902-2

Figure Lengend Snippet: Role of p38 MAPK in the expression of LOC105375913 in tubular cells. ( a ) RT-PCR analysis of LOC105375913 in HK-2 cells treated with C3a, SB203580, PD098059 and MK2206 (n = 5); ( b ) RNA pull-down and RT-PCR analysis of the binding between LOC105375913 and miR-27b in HK-2 cells treated with C3a and SB203580 (n = 5); ( c ) Western blot analysis of snail, FN and Col I in HK-2 cells treated with C3a and SB203580 (n = 3); ( d,e ) Western blot analysis of p-p38 in HK-2 cells treated with C3a or plenti-CMV-LOC105375913 plasmid (n = 3); ( f ) RT-PCR analysis of LOC105375913 in HK-2 cells treated with U46619 (n = 5); ( g ) RNA pull-down and RT-PCR analysis of the binding between LOC105375913 and miR-27b in HK-2 cells treated with U46619 (n = 5); ( h ) Western blot analysis of snail, FN and Col I in HK-2 cells treated with U46619 (n = 3). For statistical analysis, one-way ANOVA with Tukey’s post hoc test was used for ( a , b and g ) and a two-tailed Student’s t-test was used for ( f ). * P < 0.05 compared with control. # P < 0.05 compared with C3a-treated cells.

Article Snippet: For intervention studies, 1 μM of C3aR antagonist SB290157 (sc-222291, Santa Cruz), 100 μg/ml of eculizumab (Soliris, Alexion Pharmaceuticals), 10 μM of p38 MAPK inhibitor SB203580 (sc-3533, Santa Cruz), 50 μM of ERK inhibitor PD98059 (sc-3532, Santa Cruz), 10 μM of Akt inhibitor MK2206 (sc-364537, Santa Cruz) or 1 μM of U-46619 (sc-201242, Santa Cruz) was given 30 min before treatments.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Binding Assay, Western Blot, Plasmid Preparation, Two Tailed Test, Control

Journal: Scientific Reports

Article Title: Upregulated long noncoding RNA LOC105375913 induces tubulointerstitial fibrosis in focal segmental glomerulosclerosis

doi: 10.1038/s41598-018-36902-2

Figure Lengend Snippet: Role of XBP-1s in the expression of LOC105375913 in tubular cells. ( a ) RT-PCR analysis of LOC105375913 in HK-2 cells treated with C3a, XBP-1s siRNA, C/EBPβ siRNA, Elk-1 siRNA, ERα siRNA and GR siRNA (n = 3); ( b ) Level of XBP-1s and p-XBP-1s protein in HK-2 cells treated with C3a and SB203580 (n = 3); ( c ) ChIP analysis of the binding between XBP-1s and LOC105375913 promoter in HK-2 cells treated with C3a (n = 3); ( d ) Schematic of the constructed LOC105375913 promoter-luciferase reporter plasmids; ( e ) Normalized luciferase activity of reporter constructs in HK-2 cells cotransfected with XBP-1s plasmid (n = 5); ( f ) Level of LOC105375913 in HK-2 cells transfected with XBP-1s plasmid (n = 5); ( g ) RNA pull-down and RT-PCR analysis of the binding between LOC105375913 and miR-27b in HK-2 cells transfected with XBP-1s plasmid (n = 5); ( h ) Western blot analysis of snail, FN and Col I in HK-2 cells transfected with XBP-1s plasmid (n = 3); ( i ) RNA pull-down and RT-PCR analysis of the binding between LOC105375913 and miR-27b in HK-2 cells treated with C3a and XBP-1s siRNA (n = 5); ( j ) Western blot analysis of snail, FN and Col I in HK-2 cells treated with C3a and XBP-1s siRNA (n = 3). For statistical analysis, one-way ANOVA with Tukey’s post hoc test was used for ( a , e , g and i ), and a two-tailed Student’s t-test was used for ( f ). * P < 0.05 compared with control; # P < 0.05 compared with C3a-treated cells.

Article Snippet: For intervention studies, 1 μM of C3aR antagonist SB290157 (sc-222291, Santa Cruz), 100 μg/ml of eculizumab (Soliris, Alexion Pharmaceuticals), 10 μM of p38 MAPK inhibitor SB203580 (sc-3533, Santa Cruz), 50 μM of ERK inhibitor PD98059 (sc-3532, Santa Cruz), 10 μM of Akt inhibitor MK2206 (sc-364537, Santa Cruz) or 1 μM of U-46619 (sc-201242, Santa Cruz) was given 30 min before treatments.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Binding Assay, Construct, Luciferase, Activity Assay, Plasmid Preparation, Transfection, Western Blot, Two Tailed Test, Control