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: ACS Nano

Article Title: Small Gold Nanoparticles Alleviate Huntington’s Disease via Modulating p38α Mitogen-Activated Protein Kinase and Pyruvate Dehydrogenase Kinase 1

doi: 10.1021/acsnano.5c14751

Figure Lengend Snippet: Au 3 @PEG 1k NPs inhibited p38α phosphorylation and PDK1 activity in the brain of R6/2 mice. (A) Profiling 281 kinases by the Z-lyte assay revealed that ∼200 nM Au 3 @PEG 1k NPs inhibited 7 HD-related kinases by >85%. Two most HD-related kinases identified were PDK1 and p38α, based on cumulative publication count as of August 2024 derived from defined keyword searches of “Huntington’s disease” in PubMed. (B–D), 10-point titration curves and IC 50 values of Au 3 @PEG 1k NP on PDK1, MAPK14 (p38α), and MAP2K6 (MKK6; upstream of p38α) respectively. (E–F), Western blot analysis of the brain at Week 11 revealed the (i) activation of p-PDHα1 (a downstream kinase of PDK1) and p-p38α in R6/2 mice (red) when compared to age-matched healthy WT littermates (blue) and (ii) inhibition of both kinases upon Au 3 @PEG 1k NP treatment of R6/2 mice (orange) when compared to untreated R6/2 mice (red) in CTX and STR. Data are from n = 4, across 1 experiment. (G–H), Quantification of the Western blot data in (E–F) for the expression of p-PDHα1 (relative to PDHα1) and p-p38α (relative to total p38α). Statistical significance was evaluated using one-way ANOVA with Tukey’s post hoc test for multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001, **** P < 0.0001. All bars and error bars represent mean ± SD.

Article Snippet: To test the therapeutic role of selected kinases, R6/2 mice received daily i.p. injections of either (i) 0.1 mL of a vehicle (10% DMSO, 25% PEG 400 and 5% dextrose) containing an inhibitor of p38α called “p38-α MAPK-IN-1” (MedChemExpress) at a dose of 2.6 mg/kg, or (ii) 0.1 mL of D5W containing an inhibitor of PDK1, sodium dichloroacetate (DCA; Sigma, 347795) at a dose of 100 mg/kg.

Techniques: Phospho-proteomics, Activity Assay, Derivative Assay, Titration, Western Blot, Activation Assay, Inhibition, Expressing

Journal: ACS Nano

Article Title: Small Gold Nanoparticles Alleviate Huntington’s Disease via Modulating p38α Mitogen-Activated Protein Kinase and Pyruvate Dehydrogenase Kinase 1

doi: 10.1021/acsnano.5c14751

Figure Lengend Snippet: Inhibition of PDK1 activities and phosphorylation of p-p38α by their respective inhibitors in the cortex (CTX) and striatum (STR) alleviated HD and reduced pyroptosis. (A) Rotarod test of untreated (red), Au 3 @PEG 1k NP-treated (orange), and dichloroacetate (DCA; an inhibitor of PDK1)-treated (cyan) R6/2 mice as a function of age, based on the treatment plan in A. At Week 9, DCA treatments showed improved motor activity. At Weeks 11, both Au 3 @PEG 1k NP- and DCA treatments improved motor activity than untreated. Data are from n = 4, across four experiments. Statistical significance was evaluated using Two-Way ANOVA with Tukey’s post hoc test for multiple comparisons. (B) Rotarod test of untreated (red), Au 3 @PEG 1k NP-treated (orange), and p38α MAPK-IN-1 (an inhibitor of p38α)-treated (brown) R6/2 mice as a function of age, based on the treatment plan in A. At Week 7 and 9, Au 3 @PEG 1k NP-treated R6/2 mice showed improved motor activity. At Weeks 11, both Au 3 @PEG 1k NP- and p38α MAPK-IN-1 treatments improved motor activity more than untreated. Data are from n = 4, across four experiments. Statistical significance was evaluated using Two-Way ANOVA with Tukey’s post hoc test for multiple comparisons. (C–H) Western blot analysis revealed the (i) activation of p-PDHα1 (a downstream kinase of PDK1) in R6/2 mice (red) when compared to healthy WT littermates (blue) and (ii) inhibition of p-PDHα1 upon dichloroacetate (DCA; pharmacological inhibitor of PDK) treatment of R6/2 mice (cyan) when compared to untreated R6/2 mice (red) in CTX and STR. (iii) activation of p-p38α in R6/2 mice (red) when compared to healthy WT littermates (blue) and (iv) inhibition of p-p38α upon p38α MAPK-IN-1 (pharmacological inhibitor of p38α) treatment of R6/2 mice (brown) when compared to untreated R6/2 mice (red) in CTX and STR. All data are from n = 4, across one experiment. Statistical significance was evaluated using One-Way ANOVA with Tukey’s post hoc test for multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001. All bars and error bars represent mean ± SD.

Article Snippet: To test the therapeutic role of selected kinases, R6/2 mice received daily i.p. injections of either (i) 0.1 mL of a vehicle (10% DMSO, 25% PEG 400 and 5% dextrose) containing an inhibitor of p38α called “p38-α MAPK-IN-1” (MedChemExpress) at a dose of 2.6 mg/kg, or (ii) 0.1 mL of D5W containing an inhibitor of PDK1, sodium dichloroacetate (DCA; Sigma, 347795) at a dose of 100 mg/kg.

Techniques: Inhibition, Phospho-proteomics, Activity Assay, Western Blot, Activation Assay

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