Journal: Frontiers in Pharmacology

Article Title: Andrographolide Protects against Aortic Banding-Induced Experimental Cardiac Hypertrophy by Inhibiting MAPKs Signaling

doi: 10.3389/fphar.2017.00808

Figure Lengend Snippet: Andr blocks MAPKs signaling in vivo and in vitro . (A) Representative blots of phosphorylated (P-) and total (T-) ERK1/2, JNK, and P38 in the heart tissues of mice in the indicated groups ( n = 6). (B) Comparison of expression among the indicated groups. ∗ P < 0.05 compared with the corresponding sham group. # P < 0.05 vs. the veh-AB group. (C) Representative blots of phosphorylated (P-) and total (T-) ERK1/2, JNK, and P38 in the cardiomyocytes in the indicated groups ( n = 6). (D) Comparison of expression among the indicated groups. ∗ P < 0.05 compared with the corresponding PBS group. # P < 0.05 vs. the vehicle-Ang II group.

Article Snippet: Cardiomyocytes were treated with ERK1/2 inhibitor (SCH772984, 5 μM, Selleck), JNK inhibitor (SP600125, 10 μM, Sigma), and/or P38 inhibitor (SB209063, 10 μM, Medchem Express) as well as stimulated with Ang II.

Techniques: In Vivo, In Vitro, Comparison, Expressing

Journal: Frontiers in Pharmacology

Article Title: Andrographolide Protects against Aortic Banding-Induced Experimental Cardiac Hypertrophy by Inhibiting MAPKs Signaling

doi: 10.3389/fphar.2017.00808

Figure Lengend Snippet: Andr-mediated cardioprotection depends on the inhibition of MAPKs in cardiomyocytes. Cardiomyocytes were treated with ERK1/2 inhibitor (SCH7729, 5 μM), JNK inhibitor (SP600125, 10 μM), and/or P38 inhibitor (SB209063, 10 μM) as well as stimulated with Ang II (1 μM) and treated with Andr (50 μM). (A) Cell viability was accessed by the cell counting kit-8 assay ( n = 5). (B–E) Immunofluorescence staining of α-actinin and the cell surface area of cardiomyocytes in the indicated groups ( n = 5 samples and n = 100+ cells per group). (F–J) The mRNA levels of ANP and β-MHC in cardiomyocytes in the indicated groups ( n = 6). The results are presented as a fold change, and the results are normalized to GAPDH gene expression. ∗ P < 0.05 compared with the control group. # P < 0.05 vs. the Ang II group.

Article Snippet: Cardiomyocytes were treated with ERK1/2 inhibitor (SCH772984, 5 μM, Selleck), JNK inhibitor (SP600125, 10 μM, Sigma), and/or P38 inhibitor (SB209063, 10 μM, Medchem Express) as well as stimulated with Ang II.

Techniques: Inhibition, Cell Counting, Immunofluorescence, Staining, Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Andrographolide Protects against Aortic Banding-Induced Experimental Cardiac Hypertrophy by Inhibiting MAPKs Signaling

doi: 10.3389/fphar.2017.00808

Figure Lengend Snippet: Andr reduces cardiac fibroblast activation via MAPKs. (A–F) Cardiac fibroblasts were treated with different concentrations of Andr (0, 12.5, 25, or 50 μM) and/or stimulated with Ang II (1 μM). (A) Cell viability was accessed by the cell counting kit-8 assay ( n = 5). (B) Cell proliferation was accessed by the cell counting kit-8 assay ( n = 5). (C) Immunofluorescence staining of α-SMA in the indicated groups. (D) The mRNA levels of collagen I, collagen III, and CTGF in cardiac fibroblasts in the indicated groups ( n = 6). (E) Representative blots of phosphorylated (P-) and total (T-) ERK1/2, JNK, P38, and smad4 in the cardiac fibroblasts in the indicated groups ( n = 6). (F) Comparison of expression among the indicated groups. ∗ P < 0.05 compared with the corresponding PBS group. # P < 0.05 vs. the vehicle-Ang II group. (G–J) Cardiac fibroblasts were treated with ERK1/2 inhibitor (SCH7729, 5 μM), JNK inhibitor (SP600125, 10 μM), and/or P38 inhibitor (SB209063, 10 μM) as well as stimulated with Ang II (1 μM) and treated with Andr (50 μM). (G) Cell viability was accessed by the cell counting kit-8 assay ( n = 5). (H) Cell proliferation was accessed by the cell counting kit-8 assay ( n = 5). (I) Immunofluorescence staining of α-SMA in the indicated groups. (J,K) The mRNA levels of collagen I, collagen III, and CTGF in cardiac fibroblasts in the indicated groups ( n = 6). The results are presented as a fold change, and the results are normalized to GAPDH gene expression. ∗ P < 0.05 compared with the control group. # P < 0.05 vs. the Ang II group.

Article Snippet: Cardiomyocytes were treated with ERK1/2 inhibitor (SCH772984, 5 μM, Selleck), JNK inhibitor (SP600125, 10 μM, Sigma), and/or P38 inhibitor (SB209063, 10 μM, Medchem Express) as well as stimulated with Ang II.

Techniques: Activation Assay, Cell Counting, Immunofluorescence, Staining, Comparison, Expressing, Control

Journal: Oncotarget

Article Title: The oncogenic receptor ErbB2 modulates gemcitabine and irinotecan/SN-38 chemoresistance of human pancreatic cancer cells via hCNT1 transporter and multidrug-resistance associated protein MRP-2.

doi: 10.18632/oncotarget.3414

Figure Lengend Snippet: Figure 5: Identification of the signaling pathways involved in resistance to SN-38 in ErbB2-KD cells and MRP2 expression regulation. A. Western blots were performed to analyze the expression and phosphorylation of ERK1/2, JNK, p38, NF-κB, Akt and β-actin in CAPAN-2 NT and ErbB2-KD cells. The density of each marker was measured and phosphorylated/constitutive or protein/β-actin ratio was determined and represented as histograms. Expression in NT cells was arbitrarily set to 1. Three independent experiments were performed. B. Cell survival rate was measured following Erk1/2, JNK and NF-κB inhibition in CAPAN-1 and CAPAN-2 cells using specific siRNA during 48 h before SN-38 treatment. C. CAPAN-1 and CAPAN-2 cells were transiently transfected with ErbB2, Erk1/2, JNK, NF-κB or control siRNA for 48 h. qPCR were performed to analyze the expression of MRP2.

Article Snippet: Transient inhibitions of ABCG2 (sc-41151), MRP1 (sc-35962) MRP2 (sc-35963), Akt (sc-43609) and p38 (sc-29433) were performed using a pool of siRNA designed by Santa Cruz Biotechnology following cell transfection with Effectene® (Qiagen) as described by the manufacturer.

Techniques: Protein-Protein interactions, Expressing, Western Blot, Phospho-proteomics, Marker, Inhibition, Transfection, Control

Journal: Molecular medicine reports

Article Title: Ligustrazine attenuates the platelet-derived growth factor-BB-induced proliferation and migration of vascular smooth muscle cells by interrupting extracellular signal-regulated kinase and P38 mitogen-activated protein kinase pathways.

doi: 10.3892/mmr.2015.3383

Figure Lengend Snippet: Figure 5. Ligustrazine suppresses PDGF‑BB‑stimulated activation of ERK and p38 MAPK signaling in VSMCs. Western blotting was used to determine the activity of ERK and p38 MAPK signaling in each group. Control VSMCs were cultured for 48 h without any treatment. PDGF‑BB VSMCs were treated only with PDGF‑BB for 48 h. PDGF‑BB+ligustrazine VSMCs were treated with ligustrazine and PDGF‑BB for 48 h. **P<0.01, vs. control. VSMC, vascular smooth muscle cell; PDGF, platelet‑derived growth factor; ERK, extracellular signal‑related kinase; MAPK, mitogen‑activated protein kinase.

Article Snippet: Mouse monoclonal anti-desmin (1:100; sc-365130), anti-smoothelin (1:50; sc-376902), anti-phosphorylated (phosph)-ERK (1:100; sc-136521), anti-total ERK (1:50; sc-135900), anti-phospho-p38 (1:50; sc-81621), anti-total p38 (1:100; sc-7973), anti-cyclin-dependent kinase (CDK)2 (1:100; sc-6248), anti-CDK4 (1:50; sc-23896), anti-cyclin D1 (1:50; sc-450), anti-cyclin E (1:100; sc-48420) and anti-glyceraldehyde 3-phosphate dehy drogenase (GAPDH; 1:50; sc-365062) primary antibodies as well as rabbit anti-mouse horseradish peroxidase-conjugated secondary antibodies (1:5,000; sc-358914) were obtained from Santa Cruz Biotechnology, Inc., (Santa Cruz, CA, USA).

Techniques: Activation Assay, Western Blot, Activity Assay, Control, Cell Culture

Journal: The Journal of Biological Chemistry

Article Title: Knocking out p38α+p38β+p38γ is required to abort the myogenic program in C2C12 myoblasts and to impose uncontrolled proliferation

doi: 10.1016/j.jbc.2025.108281

Figure Lengend Snippet: C2p38α −/− cells are capable of executing myoblast to myotube differentiation and of expression of myogenic markers, perhaps through phosphorylation of other p38 isoforms. C2C12 and C2p38α −/− cells were cultured in growth medium (GM) to subconfluency. The medium was then changed to differentiation medium (DM) for additional 7 days. A , cells were photographed at the indicated days. B , protein lysates were prepared at the indicated time points. Twenty micrograms of protein lysates were separated by SDS-PAGE, blotted, and probed with the indicated antibodies. C , cells were fixed at day 7, immune-stained with anti MyHC antibody, and also stained with DAPI. DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: C2p38α −/− cells were produced using a p38α CRISPR/Cas9 KO plasmid (Santa Cruz, sc-424051).

Techniques: Expressing, Phospho-proteomics, Cell Culture, SDS Page, Staining

Journal: The Journal of Biological Chemistry

Article Title: Knocking out p38α+p38β+p38γ is required to abort the myogenic program in C2C12 myoblasts and to impose uncontrolled proliferation

doi: 10.1016/j.jbc.2025.108281

Figure Lengend Snippet: A pan-p38 inhibitor, as well as a p38α/β-specific inhibitor, impairs differentiation of C2p38α −/− cells. C2p38α −/− cells were cultured in growth medium to subconfluency. The medium was then changed to differentiation medium (DM) for additional 5 days. The DM was supplemented with either DMSO, 1 μM of BIRB796, or 10 μM of SB203580. A , cells were photographed at the indicated days following beginning of treatment. B , lysates were prepared at the indicated time points. Twenty micrograms of protein lysates were separated by SDS-PAGE, blotted, and probed with the indicated antibodies. DMSO, dimethyl sulfoxide.

Article Snippet: C2p38α −/− cells were produced using a p38α CRISPR/Cas9 KO plasmid (Santa Cruz, sc-424051).

Techniques: Cell Culture, SDS Page

Journal: The Journal of Biological Chemistry

Article Title: Knocking out p38α+p38β+p38γ is required to abort the myogenic program in C2C12 myoblasts and to impose uncontrolled proliferation

doi: 10.1016/j.jbc.2025.108281

Figure Lengend Snippet: p38α or p38β is required to induce expression of myomerger and myomaker in differentiating C2C12. Cells were cultured in growth medium (GM) to subconfluency. The medium was then changed to differentiation medium (DM) for additional 7 days. A and B , protein lysates prepared from C2C12, C2p38β −/− , and C2p38α/β −/− cells (A) and from C2C12 and C2p38α/β/γ −/− cells (B) were tested with the indicated antibodies. C , parental C2C12 and the indicated C2p38KO cells were incubated in DM for 5 days. Lysates were prepared and analyzed by Western blots with the indicated antibodies. Note that the GAPDH panel is the same as in D and E , as the Western blots shown in these figures were performed together on the same lysates.

Article Snippet: C2p38α −/− cells were produced using a p38α CRISPR/Cas9 KO plasmid (Santa Cruz, sc-424051).

Techniques: Expressing, Cell Culture, Incubation, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Knocking out p38α+p38β+p38γ is required to abort the myogenic program in C2C12 myoblasts and to impose uncontrolled proliferation

doi: 10.1016/j.jbc.2025.108281

Figure Lengend Snippet: C2C12 cells respond to p38 inhibitors by elevating levels of MKK6 and phosphorylation of p38 isoforms. Some myotubes do appear following 5 days of incubation with inhibitors. A , the myogenic program is functional in the presence of SB203580 and BIRB796. C2C12 cells were incubated for 5 days with either DM, DM + DMSO, or DM+ the indicated inhibitor, with addition of fresh media every 24 h. Protein lysates were prepared and analyzed by western blot with the indicated antibodies. B , the myoblast to myotube process of C2C12 cells is delayed and weakened, but not totally abolished. C2C12 were treated as described in ( A ) and photographed daily. DM, differentiation medium.

Article Snippet: C2p38α −/− cells were produced using a p38α CRISPR/Cas9 KO plasmid (Santa Cruz, sc-424051).

Techniques: Phospho-proteomics, Incubation, Functional Assay, Western Blot

Journal: Scientific Reports

Article Title: Bafilomycin A1 induces caspase-independent cell death in hepatocellular carcinoma cells via targeting of autophagy and MAPK pathways

doi: 10.1038/srep37052

Figure Lengend Snippet: ( A ) BEL7402 and HepG2 cells were treated with vehicle controls or with 5 nM BafA1 for 24, 48 and 72 h. IB analysis for p-ERK1/2, p-JNK, p-p38, total-ERK1/2, total-JNK, and total-p38 was used to examine protein expression, using β-actin as a loading control. ( B ) HCC cells were pre-treated with inhibitors to either ERK (PD98059), JNK (SP600125), or p38 (SB202190) and subsequently treated with BafA1 alone, or in combination, for 24 h, double stained with Annexin V/PI and analyzed by FACS. ( C ) HCC cells were transfected with a stable knockdown for p38 (BEL7402/shp38 and HepG2/shp38), or control cells (BEL7402/shControl and HepG2/shControl) and were treated with vehicle control or 5 nM BafA1 for 24, 48, and 72 h. At each time-point, cell viability was examined using the MTT assay. ( D ) BEL7402 and HepG2 cells were pre-treated SB202190, vehicle control or 5 nM BafA1 for 24 h. IB analysis for protein expression of Puma was carried out, using β-actin as a loading control. ( E ) BEL7402 and HepG2 cells were infected with adenoviruses expressing GFP-Puma or vector control at a multiplicity of infection of 250. Cells were treated with BafA1 and cell viability determined using the MTT assay. Data are presented as mean ± SEM from three independent experiments, (BafA1 means Bafilomycin A1).

Article Snippet: The following lentiviral constructs were purchased from Santa Cruz: MAP LC3β shRNA (sc-43390-V), p38α shRNA (sc-29433-V) and noncoding shRNA (sc-108080).

Techniques: Expressing, Control, Staining, Transfection, Knockdown, MTT Assay, Infection, Plasmid Preparation

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Puerarin Attenuates Cardiac Hypertrophy Partly Through Increasing Mir-15b/195 Expression and Suppressing Non-Canonical Transforming Growth Factor Beta (Tgfβ) Signal Pathway

doi: 10.12659/MSM.895877

Figure Lengend Snippet: Primer sequences for qRT-PCR.

Article Snippet: The ready-to-use Smad2/3 and Smad4 shRNA lentiviral particles (sc-37239-V and sc-29485-V) and p38 shRNA lentiviral particles (sc-29434-V) were purchased from Santa Cruz Biotech (Santa Cruz, CA, USA).

Techniques:

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Puerarin Attenuates Cardiac Hypertrophy Partly Through Increasing Mir-15b/195 Expression and Suppressing Non-Canonical Transforming Growth Factor Beta (Tgfβ) Signal Pathway

doi: 10.12659/MSM.895877

Figure Lengend Snippet: Puerarin suppresses the canonical and non-canonical TGFβ signal pathways partially through miR-15b and miR-195. ( A ) QRT-PCR analysis of the TGFβ receptors (TGFBR1, TGFBR2 and TGFBR3), canonical (Smad2, Smad3, Smad4, and Smad7) and non-canonical TGFβ signal members (TAK1 and p38) in the ventricular tissues in the sham, Ang II-infused, and puerarin groups. ( B ) Western blot analysis of Smad2, Smad3, Smad4, Smad7, TAK1, and p38 expression in the groups indicated in Figure A. ( C ) QRT-PCR analysis of miR-15 family expression in the primary cardiomyocytes after treatment with puerarin. ( D, E ) QRT-PCR analysis of miR-15b and miR-195 expression in the primary cardiomyocytes after infection with the pLV-miR-15b and pLV-miR-195 expression ( D ) or the pLV-miR-15b and pLV-miR-195 locker lentiviral particles ( E ). ( F, G, H ) QRT-PCR analysis Smad2, Smad3, Smad4, Smad7, and p38 expression in the primary cardiomyocytes with miR-15b or miR-195 overexpression ( F ) or miR-15b or miR-195 knockdown ( G ) or after treatment with puerarin with or without knockdown of endogenous miR-15b and miR-195 ( H ). * Comparison with NC group; # comparison with puerarin group. * and # p<0.05, ** and ## p<0.01, *** and ### p<0.001.

Article Snippet: The ready-to-use Smad2/3 and Smad4 shRNA lentiviral particles (sc-37239-V and sc-29485-V) and p38 shRNA lentiviral particles (sc-29434-V) were purchased from Santa Cruz Biotech (Santa Cruz, CA, USA).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Infection, Over Expression, Knockdown, Comparison

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Puerarin Attenuates Cardiac Hypertrophy Partly Through Increasing Mir-15b/195 Expression and Suppressing Non-Canonical Transforming Growth Factor Beta (Tgfβ) Signal Pathway

doi: 10.12659/MSM.895877

Figure Lengend Snippet: MiR-15b attenuates cardiac hypertrophy through the non-canonical TGFβ pathway. ( A, B ) QRT-PCR analysis of mRNA expression of ANP, BNP, and β-MHC in the primary cardiomyocytes after knockdown of endogenous miR-15b ( A ) or knockdown of endogenous TAK1 or p38 separately ( B ). ( C–H ) Measurement of cell surface area ( C, D ), the rate of protein synthesis ( E, F ), and the total protein content ( G, H ) in the cardiomyocytes after knockdown of endogenous miR-15b ( C, E, G ) or knockdown of endogenous TAK1 or p38 separately ( D, F, H ). * Comparison with shRNA NC group; # comparison with Ang II+ shRNA NC group. * and # p<0.05, ** and ## p<0.01, *** and ### p<0.001.

Article Snippet: The ready-to-use Smad2/3 and Smad4 shRNA lentiviral particles (sc-37239-V and sc-29485-V) and p38 shRNA lentiviral particles (sc-29434-V) were purchased from Santa Cruz Biotech (Santa Cruz, CA, USA).

Techniques: Quantitative RT-PCR, Expressing, Knockdown, Comparison, shRNA

Journal: BMC Biology

Article Title: FGF-independent MEK1/2 signalling in the developing foetal testis is essential for male germline differentiation in mice

doi: 10.1186/s12915-023-01777-x

Figure Lengend Snippet: Summary of treatments and doses used in gonad cultures

Article Snippet: PH-797804 (p38i) , 500 nM , IC50 – p38α: 26 nM; p38β 102 nM , Phase II, NCT00559910 , MedChem Express, HY-10403 , [ ] .

Techniques: Control, Recombinant