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phospho p38 mitogen activated protein kinase thr180 tyr182  (Cell Signaling Technology Inc)
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Cell Signaling Technology Inc phospho p38 mitogen activated protein kinase thr180 tyr182
NaHS and DATS protect against ACR-induced urothelial cell injury. (A,B) Effect of DATS on H 2 S production capacity in urothelial cells. SV-HUC-1 cells treated with indicated concentrations of DATS were subjected to lead sulfide assay for 24 h (A) . Densitometric quantification of the intensity of the colored circles was performed, and data shown in (B) have been normalized to control (mean ± S.E.; n = 5; ** P < 0.01). (C,D) Effect of NaHS and DATS on ACR-induced urothelial cell death. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 100 μM ACR for an additional 24 h. The cell viability was determined by Calcein-AM/PI staining (upper) and WST assay (lower). Data shown are mean ± S.E. (n = 5 and 6, respectively; ** P < 0.01). (E,F) Effect of NaHS and DATS on ACR-induced oxidative stress in urothelial cells. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 75 μM ACR for an additional 4 h. Cell proteins were extracted for Western blot analysis of protein carbonylation and phosphorylated <t>P38MAPK.</t> A quantitative analysis of blots is shown below. Data are presented as mean ± S.E. (n = 3; * P < 0.05, ** P < 0.01).
Phospho P38 Mitogen Activated Protein Kinase Thr180 Tyr182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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phospho protein 38 p p38  (Cell Signaling Technology Inc)
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Cell Signaling Technology Inc phospho protein 38 p p38
NaHS and DATS protect against ACR-induced urothelial cell injury. (A,B) Effect of DATS on H 2 S production capacity in urothelial cells. SV-HUC-1 cells treated with indicated concentrations of DATS were subjected to lead sulfide assay for 24 h (A) . Densitometric quantification of the intensity of the colored circles was performed, and data shown in (B) have been normalized to control (mean ± S.E.; n = 5; ** P < 0.01). (C,D) Effect of NaHS and DATS on ACR-induced urothelial cell death. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 100 μM ACR for an additional 24 h. The cell viability was determined by Calcein-AM/PI staining (upper) and WST assay (lower). Data shown are mean ± S.E. (n = 5 and 6, respectively; ** P < 0.01). (E,F) Effect of NaHS and DATS on ACR-induced oxidative stress in urothelial cells. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 75 μM ACR for an additional 4 h. Cell proteins were extracted for Western blot analysis of protein carbonylation and phosphorylated <t>P38MAPK.</t> A quantitative analysis of blots is shown below. Data are presented as mean ± S.E. (n = 3; * P < 0.05, ** P < 0.01).
Phospho Protein 38 P P38, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti phospho p38 mitogen activated protein kinase mapk rabbit monoclonal antibody  (Cell Signaling Technology Inc)
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NaHS and DATS protect against ACR-induced urothelial cell injury. (A,B) Effect of DATS on H 2 S production capacity in urothelial cells. SV-HUC-1 cells treated with indicated concentrations of DATS were subjected to lead sulfide assay for 24 h (A) . Densitometric quantification of the intensity of the colored circles was performed, and data shown in (B) have been normalized to control (mean ± S.E.; n = 5; ** P < 0.01). (C,D) Effect of NaHS and DATS on ACR-induced urothelial cell death. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 100 μM ACR for an additional 24 h. The cell viability was determined by Calcein-AM/PI staining (upper) and WST assay (lower). Data shown are mean ± S.E. (n = 5 and 6, respectively; ** P < 0.01). (E,F) Effect of NaHS and DATS on ACR-induced oxidative stress in urothelial cells. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 75 μM ACR for an additional 4 h. Cell proteins were extracted for Western blot analysis of protein carbonylation and phosphorylated <t>P38MAPK.</t> A quantitative analysis of blots is shown below. Data are presented as mean ± S.E. (n = 3; * P < 0.05, ** P < 0.01).
Anti Phospho P38 Mitogen Activated Protein Kinase Mapk Rabbit Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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instantonetm elisa kit for p38 mitogen-activated protein kinase (p38-mapk) (phospho) [pt180/py182]  (Thermo Fisher)
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NaHS and DATS protect against ACR-induced urothelial cell injury. (A,B) Effect of DATS on H 2 S production capacity in urothelial cells. SV-HUC-1 cells treated with indicated concentrations of DATS were subjected to lead sulfide assay for 24 h (A) . Densitometric quantification of the intensity of the colored circles was performed, and data shown in (B) have been normalized to control (mean ± S.E.; n = 5; ** P < 0.01). (C,D) Effect of NaHS and DATS on ACR-induced urothelial cell death. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 100 μM ACR for an additional 24 h. The cell viability was determined by Calcein-AM/PI staining (upper) and WST assay (lower). Data shown are mean ± S.E. (n = 5 and 6, respectively; ** P < 0.01). (E,F) Effect of NaHS and DATS on ACR-induced oxidative stress in urothelial cells. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 75 μM ACR for an additional 4 h. Cell proteins were extracted for Western blot analysis of protein carbonylation and phosphorylated <t>P38MAPK.</t> A quantitative analysis of blots is shown below. Data are presented as mean ± S.E. (n = 3; * P < 0.05, ** P < 0.01).
Instantonetm Elisa Kit For P38 Mitogen Activated Protein Kinase (P38 Mapk) (Phospho) [Pt180/Py182], supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti-phospho-p38 mitogen-activated protein kinase (p38 mapk) (thr180/tyr182) er2001-52  (Huabio Inc)
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NaHS and DATS protect against ACR-induced urothelial cell injury. (A,B) Effect of DATS on H 2 S production capacity in urothelial cells. SV-HUC-1 cells treated with indicated concentrations of DATS were subjected to lead sulfide assay for 24 h (A) . Densitometric quantification of the intensity of the colored circles was performed, and data shown in (B) have been normalized to control (mean ± S.E.; n = 5; ** P < 0.01). (C,D) Effect of NaHS and DATS on ACR-induced urothelial cell death. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 100 μM ACR for an additional 24 h. The cell viability was determined by Calcein-AM/PI staining (upper) and WST assay (lower). Data shown are mean ± S.E. (n = 5 and 6, respectively; ** P < 0.01). (E,F) Effect of NaHS and DATS on ACR-induced oxidative stress in urothelial cells. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 75 μM ACR for an additional 4 h. Cell proteins were extracted for Western blot analysis of protein carbonylation and phosphorylated <t>P38MAPK.</t> A quantitative analysis of blots is shown below. Data are presented as mean ± S.E. (n = 3; * P < 0.05, ** P < 0.01).
Anti Phospho P38 Mitogen Activated Protein Kinase (P38 Mapk) (Thr180/Tyr182) Er2001 52, supplied by Huabio Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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phospho p38 mitogen activated protein kinase mapk  (Proteintech)
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Proteintech phospho p38 mitogen activated protein kinase mapk
EPHB1‐EFNB1 mediates platelet activation via the phosphorylation of AKT. (A) The tdTomato‐positive platelets isolated from Rosa26 mT/mG mice were preincubated with EphB1 or EFNB1 antibodies alone and then mixed with tumor cells. Tumor cells without platelets were used as controls. Representative flow cytometry analysis of tumor cells aggregated with tdTomato‐positive platelets. (B) Graph showing the percentage of tdTomato‐positive tumor cells that were aggregated with platelets. Data were presented as the means ± SD, with n = 3 per group. (C) Platelets were treated with Fc fragment, thrombin, EphB1‐Fc, thrombin + EphB1‐Fc, mono EphB1, mono EphB1 + EphB1‐Fc, or EFNB1 Ab + EphB1‐Fc. Platelets without treatment served as controls. CD62P expression in activated platelets was tested by flow cytometry analysis. Representative flow cytometry analysis of CD62P expression is presented on the left. The right graph shows the percentage of CD62P + platelets. Data were presented as the means ± SD, with n = 3 per group. (D) Western blot analysis showing p‐AKT (Ser473) and MAPK pathway protein levels of <t>p‐P38,</t> p‐JNK and p‐ERK after treatment with EphB1‐Fc or thrombin. (E‐F) Platelets were treated with or without LY294002 (10 µmol/L) for 20 min and then treated with EphB1‐Fc for another 20 min. CD62P expression in platelets was tested by flow cytometry analysis. A representative flow cytometry analysis of CD62P expression is presented in (E). The graph in (F) shows the percentage of CD62P + platelets. Data were presented as the means ± SD, with n = 3 per group. Statistical analyses were performed using one‐way ANOVA followed by Tukey's multiple comparisons test in (B, C, F). * P <0.05, ** P <0.01, *** P <0.001. Abbreviations: AKT, Protein kinase B; mono EphB1, EphB1 monomers, monomeric exodomains of EPHB1; EphB1‐Fc, Recombinant EphB1 Fc chimera protein; MAPK, Mitogen‐activated protein kinase; JNK, c‐Jun N‐terminal kinase; ERK, Extracellular signal‐regulated kinase; One‐way ANOVA, One‐way analysis of variance; SD, Standard deviations.
Phospho P38 Mitogen Activated Protein Kinase Mapk, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti-phospho p38 mitogen-activated protein kinase (p-p38 mapk) alpha thr180/tyr182 antibody  (Thermo Fisher)
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Thermo Fisher anti-phospho p38 mitogen-activated protein kinase (p-p38 mapk) alpha thr180/tyr182 antibody
Microarray analysis reveals that <t>p38</t> MAPK signaling pathway is involved in CPT1 inhibition induces cardiomyocytes proliferation. (A) Heat map of differentially expressed genes in hearts from ETX-treated or control mice. (B) The results of GO analysis of up- or down-regulated genes. (C) ETX down-regulated fatty acid metabolism regulating genes, including acyl-Coenzyme A dehydrogenase, short chain ( Acads ), acyl-Coenzyme A dehydrogenase, short/branched chain ( Acadsb ), acyl-CoA synthetase long chain family member 5 ( Acsl5 ) and fatty acid transporter, member 1 ( Fat1p1 ) ( n = 3, ∗ P < 0.05 vs. Control). (D) ETX up-regulated glycolysis regulating genes, including Gck , 2,3-bisphosphoglycerate mutase ( Bpgm ), and phosphofructokinase-1 ( Pfk1 ) ( n = 3, ∗ P < 0.05 vs. Control). (E) Cyclin A2 ( Ccna2 ), cyclin B1 ( Ccnb1 ), cyclin B2 ( Ccnb2 ), cyclin E1 ( Ccne1 ), cyclin-dependent kinase 1 ( Cdk1 ), CDC28 protein kinase regulatory subunit 2 ( Cks2 ), cell division cycle associated 8 ( Cdca8 ), cell division cycle 25C ( Cdc25c ), polo-like kinase 1 ( Plk1 ), cyclin-dependent kinase inhibitor 1B ( Cdkn1b ), and CDK5 regulatory subunit associated protein 1 ( Cdk5rap1 ) gene expression with ETX or without (Control) ETX treatment ( n = 3, ∗ P < 0.05 vs. control). (F) Heat map of differentially expressed genes related to p38 MAPK signaling after ETX treatment. Error bars indicate SEM.
Anti Phospho P38 Mitogen Activated Protein Kinase (P P38 Mapk) Alpha Thr180/Tyr182 Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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protein 1 light chain 3 lc3  (Cell Signaling Technology Inc)
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Fig. 11. Endogenous tau levels are decreased following treatment with the Syk inhibitor BAY 61–3606. Endogenous total tau (Tau5) and phosphorylated tau (PHF-1) levels are decreased in noninduced M1C cells following treatment with the Syk inhibitor BAY 61–3606. Data are presented as mean ± SD (N = 4). All data were normally distributed and were analyzed using Student’s t-test. *p < 0.05, **p < 0.01. Abbreviations: GAPDH, glyceraldehyde 3-phosphate dehy- drogenase; <t>LC3,</t> microtubule-associated protein 1 light chain 3; SD, standard deviation; Syk, splenic tyrosine kinase.
Protein 1 Light Chain 3 Lc3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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protein kinase pp38mapk  (Cell Signaling Technology Inc)
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Molecular Regulation of Fibrosis in Ex Vivo Cultured Hearts Representative pictures of the right ventricle and the right atrium of hearts cultured for 7 days under standard conditions (−), in the presence of SB431542 (SB, SB1, SB2) or transforming growth factor-beta (TGF-β) for (A) phospho-SMAD2/3 (pSMAD2/3) and phospho-p38 mitogen-activated protein kinase <t>(pp38MAPK),</t> (B) α-SMA and vimentin, (C) PECAM-1 and cTnI, (D) collagen I and collagen III, and (E) Alcian blue, periostin, versican B, and fibronectin. (F) Graphs depicting the percentage of hearts with α-SMA expression in the subepicardium of the ventricle and atrium after culture for 7 days under standard conditions (−), in the presence of SB431542 (SB) or TGF-β. (G and H) Graphs depicting the relative α-SMA-positive area (G) and the relative vimentin-positive area (H) in the ventricular or atrial myocardium of hearts cultured for 7 days under standard conditions (−), in the presence of SB431542 (SB) or TGF-β. DAPI was used as counterstain in all fluorescent pictures. Data are presented as means ± SEM. To evaluate significant differences, 1-way analysis of variance followed by Tukey’s multiple comparisons test was performed for graphs (G and H). Kruskal-Wallis test followed by Dunn’s multiple comparisons test was performed for graph (G [atrium]). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Scale bar: 20 μm. Abbreviations as in <xref ref-type=Figure 1 . " width="250" height="auto" />
Protein Kinase Pp38mapk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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NaHS and DATS protect against ACR-induced urothelial cell injury. (A,B) Effect of DATS on H 2 S production capacity in urothelial cells. SV-HUC-1 cells treated with indicated concentrations of DATS were subjected to lead sulfide assay for 24 h (A) . Densitometric quantification of the intensity of the colored circles was performed, and data shown in (B) have been normalized to control (mean ± S.E.; n = 5; ** P < 0.01). (C,D) Effect of NaHS and DATS on ACR-induced urothelial cell death. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 100 μM ACR for an additional 24 h. The cell viability was determined by Calcein-AM/PI staining (upper) and WST assay (lower). Data shown are mean ± S.E. (n = 5 and 6, respectively; ** P < 0.01). (E,F) Effect of NaHS and DATS on ACR-induced oxidative stress in urothelial cells. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 75 μM ACR for an additional 4 h. Cell proteins were extracted for Western blot analysis of protein carbonylation and phosphorylated P38MAPK. A quantitative analysis of blots is shown below. Data are presented as mean ± S.E. (n = 3; * P < 0.05, ** P < 0.01).

Journal: Frontiers in Pharmacology

Article Title: Hydrogen sulfide and ferroptosis inhibition underlies the dietary restriction-induced protection against cyclophosphamide cystitis

doi: 10.3389/fphar.2025.1562852

Figure Lengend Snippet: NaHS and DATS protect against ACR-induced urothelial cell injury. (A,B) Effect of DATS on H 2 S production capacity in urothelial cells. SV-HUC-1 cells treated with indicated concentrations of DATS were subjected to lead sulfide assay for 24 h (A) . Densitometric quantification of the intensity of the colored circles was performed, and data shown in (B) have been normalized to control (mean ± S.E.; n = 5; ** P < 0.01). (C,D) Effect of NaHS and DATS on ACR-induced urothelial cell death. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 100 μM ACR for an additional 24 h. The cell viability was determined by Calcein-AM/PI staining (upper) and WST assay (lower). Data shown are mean ± S.E. (n = 5 and 6, respectively; ** P < 0.01). (E,F) Effect of NaHS and DATS on ACR-induced oxidative stress in urothelial cells. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 75 μM ACR for an additional 4 h. Cell proteins were extracted for Western blot analysis of protein carbonylation and phosphorylated P38MAPK. A quantitative analysis of blots is shown below. Data are presented as mean ± S.E. (n = 3; * P < 0.05, ** P < 0.01).

Article Snippet: Antibodies against γ-H2A.X (#9718), GPX4 (#52455), phospho-p38 mitogen activated protein kinase (Thr180/Tyr182) (p-P38MAPK, #8203S), α-tubulin (#3873T), rabbit IgG (#5366), and mouse IgG (#5470) were from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Control, Staining, WST Assay, Western Blot

EPHB1‐EFNB1 mediates platelet activation via the phosphorylation of AKT. (A) The tdTomato‐positive platelets isolated from Rosa26 mT/mG mice were preincubated with EphB1 or EFNB1 antibodies alone and then mixed with tumor cells. Tumor cells without platelets were used as controls. Representative flow cytometry analysis of tumor cells aggregated with tdTomato‐positive platelets. (B) Graph showing the percentage of tdTomato‐positive tumor cells that were aggregated with platelets. Data were presented as the means ± SD, with n = 3 per group. (C) Platelets were treated with Fc fragment, thrombin, EphB1‐Fc, thrombin + EphB1‐Fc, mono EphB1, mono EphB1 + EphB1‐Fc, or EFNB1 Ab + EphB1‐Fc. Platelets without treatment served as controls. CD62P expression in activated platelets was tested by flow cytometry analysis. Representative flow cytometry analysis of CD62P expression is presented on the left. The right graph shows the percentage of CD62P + platelets. Data were presented as the means ± SD, with n = 3 per group. (D) Western blot analysis showing p‐AKT (Ser473) and MAPK pathway protein levels of p‐P38, p‐JNK and p‐ERK after treatment with EphB1‐Fc or thrombin. (E‐F) Platelets were treated with or without LY294002 (10 µmol/L) for 20 min and then treated with EphB1‐Fc for another 20 min. CD62P expression in platelets was tested by flow cytometry analysis. A representative flow cytometry analysis of CD62P expression is presented in (E). The graph in (F) shows the percentage of CD62P + platelets. Data were presented as the means ± SD, with n = 3 per group. Statistical analyses were performed using one‐way ANOVA followed by Tukey's multiple comparisons test in (B, C, F). * P <0.05, ** P <0.01, *** P <0.001. Abbreviations: AKT, Protein kinase B; mono EphB1, EphB1 monomers, monomeric exodomains of EPHB1; EphB1‐Fc, Recombinant EphB1 Fc chimera protein; MAPK, Mitogen‐activated protein kinase; JNK, c‐Jun N‐terminal kinase; ERK, Extracellular signal‐regulated kinase; One‐way ANOVA, One‐way analysis of variance; SD, Standard deviations.

Journal: Cancer Communications

Article Title: Reciprocal tumor‐platelet interaction through the EPHB1‐EFNB1 axis in the liver metastatic niche promotes metastatic tumor outgrowth in pancreatic ductal adenocarcinoma

doi: 10.1002/cac2.12637

Figure Lengend Snippet: EPHB1‐EFNB1 mediates platelet activation via the phosphorylation of AKT. (A) The tdTomato‐positive platelets isolated from Rosa26 mT/mG mice were preincubated with EphB1 or EFNB1 antibodies alone and then mixed with tumor cells. Tumor cells without platelets were used as controls. Representative flow cytometry analysis of tumor cells aggregated with tdTomato‐positive platelets. (B) Graph showing the percentage of tdTomato‐positive tumor cells that were aggregated with platelets. Data were presented as the means ± SD, with n = 3 per group. (C) Platelets were treated with Fc fragment, thrombin, EphB1‐Fc, thrombin + EphB1‐Fc, mono EphB1, mono EphB1 + EphB1‐Fc, or EFNB1 Ab + EphB1‐Fc. Platelets without treatment served as controls. CD62P expression in activated platelets was tested by flow cytometry analysis. Representative flow cytometry analysis of CD62P expression is presented on the left. The right graph shows the percentage of CD62P + platelets. Data were presented as the means ± SD, with n = 3 per group. (D) Western blot analysis showing p‐AKT (Ser473) and MAPK pathway protein levels of p‐P38, p‐JNK and p‐ERK after treatment with EphB1‐Fc or thrombin. (E‐F) Platelets were treated with or without LY294002 (10 µmol/L) for 20 min and then treated with EphB1‐Fc for another 20 min. CD62P expression in platelets was tested by flow cytometry analysis. A representative flow cytometry analysis of CD62P expression is presented in (E). The graph in (F) shows the percentage of CD62P + platelets. Data were presented as the means ± SD, with n = 3 per group. Statistical analyses were performed using one‐way ANOVA followed by Tukey's multiple comparisons test in (B, C, F). * P <0.05, ** P <0.01, *** P <0.001. Abbreviations: AKT, Protein kinase B; mono EphB1, EphB1 monomers, monomeric exodomains of EPHB1; EphB1‐Fc, Recombinant EphB1 Fc chimera protein; MAPK, Mitogen‐activated protein kinase; JNK, c‐Jun N‐terminal kinase; ERK, Extracellular signal‐regulated kinase; One‐way ANOVA, One‐way analysis of variance; SD, Standard deviations.

Article Snippet: The membranes were blocked with 5% skim milk for 1 h and then incubated overnight at 4°C with one of the following primary antibodies: EPHB1 (1:500, Abgent, or 1:500, #PA5‐14604, Invitrogen), phospho‐Akt (Ser473) (1:1,000, #4060, Cell Signaling Technology), Akt (1:1,000, #9272, Cell Signaling Technology), c‐Jun N‐terminal kinase (JNK) (1:1,000, #24164‐1‐AP, Proteintech, Wuhan, China), phospho‐JNK (Tyr185) (1:1,000, #80024‐1‐RR, Proteintech), phospho‐p38 mitogen‐activated protein kinase (MAPK) (Thr180/Tyr182) (1:1,000, #28796‐1‐AP, Proteintech), p38 MAPK (1:1,000, #14064‐1‐AP, Proteintech), p44/42 MAPK (extracellular regulated protein kinases1/2, Erk1/2) (1:1,000, #9102, Proteintech), p44/42 MAPK (Erk1/2) (137F5) (1:1,000, #4695s, Cell Signaling Technology), Phospho‐p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (1:1,000, #9101, Cell Signaling Technology) and anti‐beta‐actin (1:3,000, #AB0035, Abways, Shanghai, China).

Techniques: Activation Assay, Phospho-proteomics, Isolation, Flow Cytometry, Expressing, Western Blot, Recombinant

Microarray analysis reveals that p38 MAPK signaling pathway is involved in CPT1 inhibition induces cardiomyocytes proliferation. (A) Heat map of differentially expressed genes in hearts from ETX-treated or control mice. (B) The results of GO analysis of up- or down-regulated genes. (C) ETX down-regulated fatty acid metabolism regulating genes, including acyl-Coenzyme A dehydrogenase, short chain ( Acads ), acyl-Coenzyme A dehydrogenase, short/branched chain ( Acadsb ), acyl-CoA synthetase long chain family member 5 ( Acsl5 ) and fatty acid transporter, member 1 ( Fat1p1 ) ( n = 3, ∗ P < 0.05 vs. Control). (D) ETX up-regulated glycolysis regulating genes, including Gck , 2,3-bisphosphoglycerate mutase ( Bpgm ), and phosphofructokinase-1 ( Pfk1 ) ( n = 3, ∗ P < 0.05 vs. Control). (E) Cyclin A2 ( Ccna2 ), cyclin B1 ( Ccnb1 ), cyclin B2 ( Ccnb2 ), cyclin E1 ( Ccne1 ), cyclin-dependent kinase 1 ( Cdk1 ), CDC28 protein kinase regulatory subunit 2 ( Cks2 ), cell division cycle associated 8 ( Cdca8 ), cell division cycle 25C ( Cdc25c ), polo-like kinase 1 ( Plk1 ), cyclin-dependent kinase inhibitor 1B ( Cdkn1b ), and CDK5 regulatory subunit associated protein 1 ( Cdk5rap1 ) gene expression with ETX or without (Control) ETX treatment ( n = 3, ∗ P < 0.05 vs. control). (F) Heat map of differentially expressed genes related to p38 MAPK signaling after ETX treatment. Error bars indicate SEM.

Journal: Acta Pharmaceutica Sinica. B

Article Title: Reversing metabolic reprogramming by CPT1 inhibition with etomoxir promotes cardiomyocyte proliferation and heart regeneration via DUSP1 ADP-ribosylation-mediated p38 MAPK phosphorylation

doi: 10.1016/j.apsb.2024.11.001

Figure Lengend Snippet: Microarray analysis reveals that p38 MAPK signaling pathway is involved in CPT1 inhibition induces cardiomyocytes proliferation. (A) Heat map of differentially expressed genes in hearts from ETX-treated or control mice. (B) The results of GO analysis of up- or down-regulated genes. (C) ETX down-regulated fatty acid metabolism regulating genes, including acyl-Coenzyme A dehydrogenase, short chain ( Acads ), acyl-Coenzyme A dehydrogenase, short/branched chain ( Acadsb ), acyl-CoA synthetase long chain family member 5 ( Acsl5 ) and fatty acid transporter, member 1 ( Fat1p1 ) ( n = 3, ∗ P < 0.05 vs. Control). (D) ETX up-regulated glycolysis regulating genes, including Gck , 2,3-bisphosphoglycerate mutase ( Bpgm ), and phosphofructokinase-1 ( Pfk1 ) ( n = 3, ∗ P < 0.05 vs. Control). (E) Cyclin A2 ( Ccna2 ), cyclin B1 ( Ccnb1 ), cyclin B2 ( Ccnb2 ), cyclin E1 ( Ccne1 ), cyclin-dependent kinase 1 ( Cdk1 ), CDC28 protein kinase regulatory subunit 2 ( Cks2 ), cell division cycle associated 8 ( Cdca8 ), cell division cycle 25C ( Cdc25c ), polo-like kinase 1 ( Plk1 ), cyclin-dependent kinase inhibitor 1B ( Cdkn1b ), and CDK5 regulatory subunit associated protein 1 ( Cdk5rap1 ) gene expression with ETX or without (Control) ETX treatment ( n = 3, ∗ P < 0.05 vs. control). (F) Heat map of differentially expressed genes related to p38 MAPK signaling after ETX treatment. Error bars indicate SEM.

Article Snippet: The primary antibodies were: anti-phospho p38 mitogen-activated protein kinase (p-p38 MAPK) alpha Thr180/Tyr182 antibody (Thermo Fisher Scientific; 36-8500, Rabbit, 1:500), anti-p38 MAPK antibody (Cell Signaling Technology; 9212, Rabbit, 1:1000), anti-dual-specificity phosphatases 1 (DUSP1) antibody (Cell Signaling Technology; 48625, Rabbit, 1:1000), anti-dual-specificity phosphatases 4 (DUSP4) antibody (Cell Signaling Technology; 5149, Rabbit, 1:1000), anti-dual-specificity phosphatases 12 (DUSP12) antibody (Proteintech, Wuhan, China; 67101-1-Ig, Mouse, 1:1000), anti-Poly/Mono-ADP ribose antibody (Cell Signaling Technology; 83732, Rabbit, 1:1000), anti-poly(ADP-ribose) polymerase family, member 1 (PARP1) antibody (Proteintech; 66520-1-Ig, Mouse, 1:1000), anti-mitogen-activated protein kinase 3 (MAP2K3) antibody (Proteintech; 80137-1-RR, Rabbit, 1:1000), anti-CPT1A antibody (Abcam; ab234111, Rabbit, 1:1000), anti-CPT1B antibody (Proteintech; 22170-1-AP, Rabbit, 1:1000), and anti-glucokinase (GCK) antibody (Proteintech; 19666-1-AP, Rabbit, 1:1000).

Techniques: Microarray, Inhibition, Control, Gene Expression

CPT1 inhibition induces cardiomyocytes proliferation via p38 MAPK signaling pathway in postnatal mice. (A) The effect of ETX treatment on p38 MPAK phosphorylation in P14 heats ( n = 6, ∗ P < 0.05 vs. control). (B) The effect of ETX, p38 MAPK WT overexpression and p38 MAPK T180A/Y182A (T180A/Y182A) overexpression on p38 MPAK phosphorylation in P7 cardiomyocytes ( n = 3, ∗ P < 0.05, NS=Not significant). (C, D) The effect of ETX treatment, WT, and T180A/Y182A on the ratio of Ki67 + (red, white arrow) or PH3 + (red, white arrow) in P7 cardiomyocytes ( n = 3, ∗ P < 0.05, NS=Not significant). Scale bars = 50 μm. Error bars indicate SEM.

Journal: Acta Pharmaceutica Sinica. B

Article Title: Reversing metabolic reprogramming by CPT1 inhibition with etomoxir promotes cardiomyocyte proliferation and heart regeneration via DUSP1 ADP-ribosylation-mediated p38 MAPK phosphorylation

doi: 10.1016/j.apsb.2024.11.001

Figure Lengend Snippet: CPT1 inhibition induces cardiomyocytes proliferation via p38 MAPK signaling pathway in postnatal mice. (A) The effect of ETX treatment on p38 MPAK phosphorylation in P14 heats ( n = 6, ∗ P < 0.05 vs. control). (B) The effect of ETX, p38 MAPK WT overexpression and p38 MAPK T180A/Y182A (T180A/Y182A) overexpression on p38 MPAK phosphorylation in P7 cardiomyocytes ( n = 3, ∗ P < 0.05, NS=Not significant). (C, D) The effect of ETX treatment, WT, and T180A/Y182A on the ratio of Ki67 + (red, white arrow) or PH3 + (red, white arrow) in P7 cardiomyocytes ( n = 3, ∗ P < 0.05, NS=Not significant). Scale bars = 50 μm. Error bars indicate SEM.

Article Snippet: The primary antibodies were: anti-phospho p38 mitogen-activated protein kinase (p-p38 MAPK) alpha Thr180/Tyr182 antibody (Thermo Fisher Scientific; 36-8500, Rabbit, 1:500), anti-p38 MAPK antibody (Cell Signaling Technology; 9212, Rabbit, 1:1000), anti-dual-specificity phosphatases 1 (DUSP1) antibody (Cell Signaling Technology; 48625, Rabbit, 1:1000), anti-dual-specificity phosphatases 4 (DUSP4) antibody (Cell Signaling Technology; 5149, Rabbit, 1:1000), anti-dual-specificity phosphatases 12 (DUSP12) antibody (Proteintech, Wuhan, China; 67101-1-Ig, Mouse, 1:1000), anti-Poly/Mono-ADP ribose antibody (Cell Signaling Technology; 83732, Rabbit, 1:1000), anti-poly(ADP-ribose) polymerase family, member 1 (PARP1) antibody (Proteintech; 66520-1-Ig, Mouse, 1:1000), anti-mitogen-activated protein kinase 3 (MAP2K3) antibody (Proteintech; 80137-1-RR, Rabbit, 1:1000), anti-CPT1A antibody (Abcam; ab234111, Rabbit, 1:1000), anti-CPT1B antibody (Proteintech; 22170-1-AP, Rabbit, 1:1000), and anti-glucokinase (GCK) antibody (Proteintech; 19666-1-AP, Rabbit, 1:1000).

Techniques: Inhibition, Phospho-proteomics, Control, Over Expression

CPT1 inhibition induces cardiomyocytes proliferation via p38 MAPK signaling pathway in adult mice. (A) The effect of ETX, p38 MAPK WT overexpression and p38 MAPK T180A/Y182A (T180A/Y182A) overexpression on p38 MPAK phosphorylation in adult mice heart ( n = 3, ∗ P < 0.05, NS=Not significant). (B, C) The effect of ETX treatment, WT, and T180A/Y182A on the ratio of Ki67 + (red, white arrow) or PH3 + (red, white arrow) cardiomyocytes in adult mice hearts ( n = 5, ∗ P < 0.05, NS=Not significant; Scale bars = 20 μm). (D) LVEF, LVFS, LVIDd, LVIDs, LVEDV and LVESV at 4 weeks after MI were measured in mice after treatment with or without ETX, WT and T180A/Y182A ( n = 5, ∗ P < 0.05, NS=Not significant). (E) The infarcted area of the heart in mice 4 weeks after MI treated with or without ETX, WT and T180A/Y182A ( n = 5, ∗ P < 0.05, NS=Not significant; Scale bars = 1 mm). Error bars indicate SEM.

Journal: Acta Pharmaceutica Sinica. B

Article Title: Reversing metabolic reprogramming by CPT1 inhibition with etomoxir promotes cardiomyocyte proliferation and heart regeneration via DUSP1 ADP-ribosylation-mediated p38 MAPK phosphorylation

doi: 10.1016/j.apsb.2024.11.001

Figure Lengend Snippet: CPT1 inhibition induces cardiomyocytes proliferation via p38 MAPK signaling pathway in adult mice. (A) The effect of ETX, p38 MAPK WT overexpression and p38 MAPK T180A/Y182A (T180A/Y182A) overexpression on p38 MPAK phosphorylation in adult mice heart ( n = 3, ∗ P < 0.05, NS=Not significant). (B, C) The effect of ETX treatment, WT, and T180A/Y182A on the ratio of Ki67 + (red, white arrow) or PH3 + (red, white arrow) cardiomyocytes in adult mice hearts ( n = 5, ∗ P < 0.05, NS=Not significant; Scale bars = 20 μm). (D) LVEF, LVFS, LVIDd, LVIDs, LVEDV and LVESV at 4 weeks after MI were measured in mice after treatment with or without ETX, WT and T180A/Y182A ( n = 5, ∗ P < 0.05, NS=Not significant). (E) The infarcted area of the heart in mice 4 weeks after MI treated with or without ETX, WT and T180A/Y182A ( n = 5, ∗ P < 0.05, NS=Not significant; Scale bars = 1 mm). Error bars indicate SEM.

Article Snippet: The primary antibodies were: anti-phospho p38 mitogen-activated protein kinase (p-p38 MAPK) alpha Thr180/Tyr182 antibody (Thermo Fisher Scientific; 36-8500, Rabbit, 1:500), anti-p38 MAPK antibody (Cell Signaling Technology; 9212, Rabbit, 1:1000), anti-dual-specificity phosphatases 1 (DUSP1) antibody (Cell Signaling Technology; 48625, Rabbit, 1:1000), anti-dual-specificity phosphatases 4 (DUSP4) antibody (Cell Signaling Technology; 5149, Rabbit, 1:1000), anti-dual-specificity phosphatases 12 (DUSP12) antibody (Proteintech, Wuhan, China; 67101-1-Ig, Mouse, 1:1000), anti-Poly/Mono-ADP ribose antibody (Cell Signaling Technology; 83732, Rabbit, 1:1000), anti-poly(ADP-ribose) polymerase family, member 1 (PARP1) antibody (Proteintech; 66520-1-Ig, Mouse, 1:1000), anti-mitogen-activated protein kinase 3 (MAP2K3) antibody (Proteintech; 80137-1-RR, Rabbit, 1:1000), anti-CPT1A antibody (Abcam; ab234111, Rabbit, 1:1000), anti-CPT1B antibody (Proteintech; 22170-1-AP, Rabbit, 1:1000), and anti-glucokinase (GCK) antibody (Proteintech; 19666-1-AP, Rabbit, 1:1000).

Techniques: Inhibition, Over Expression, Phospho-proteomics

DUPS1 regulates p38 MAPK signaling after CPT1 inhibition in postnatal cardiomyocytes. (A) The expression of Map2k3 , mitogen-activated protein kinase kinase 6 ( Map2k6 ), and dual specificity phosphatase family ( Dusp1 , Dusp2 , Dusp3 , Dusp4 , Dusp5 , Dusp6 , Dusp7 , Dusp8 , Dusp9 , Dusp10 , Dusp11 , Dusp12 , Dusp13 , Dusp14 , Dusp15 , Dusp16 , Dusp18 , Dusp19 , Dusp21 , Dusp22 ) in P1 and P7 cardiomyocytes ( n = 6, ∗ P < 0.05 vs. P1). (B) The effect of ETX treatment and Dusp1 siRNA (si Dusp1 ) on DUSP1 expression and p38 MAPK phosphorylation in P7 cardiomyocytes ( n = 3, ∗ P < 0.05 vs. Control, # P < 0.05 vs. ETX). (C) The effect of ETX treatment on the interaction of DUSP1 and p38 MAPK in P7 cardiomyocytes ( n = 6, ∗ P < 0.05 vs. Control). Error bars indicate SEM.

Journal: Acta Pharmaceutica Sinica. B

Article Title: Reversing metabolic reprogramming by CPT1 inhibition with etomoxir promotes cardiomyocyte proliferation and heart regeneration via DUSP1 ADP-ribosylation-mediated p38 MAPK phosphorylation

doi: 10.1016/j.apsb.2024.11.001

Figure Lengend Snippet: DUPS1 regulates p38 MAPK signaling after CPT1 inhibition in postnatal cardiomyocytes. (A) The expression of Map2k3 , mitogen-activated protein kinase kinase 6 ( Map2k6 ), and dual specificity phosphatase family ( Dusp1 , Dusp2 , Dusp3 , Dusp4 , Dusp5 , Dusp6 , Dusp7 , Dusp8 , Dusp9 , Dusp10 , Dusp11 , Dusp12 , Dusp13 , Dusp14 , Dusp15 , Dusp16 , Dusp18 , Dusp19 , Dusp21 , Dusp22 ) in P1 and P7 cardiomyocytes ( n = 6, ∗ P < 0.05 vs. P1). (B) The effect of ETX treatment and Dusp1 siRNA (si Dusp1 ) on DUSP1 expression and p38 MAPK phosphorylation in P7 cardiomyocytes ( n = 3, ∗ P < 0.05 vs. Control, # P < 0.05 vs. ETX). (C) The effect of ETX treatment on the interaction of DUSP1 and p38 MAPK in P7 cardiomyocytes ( n = 6, ∗ P < 0.05 vs. Control). Error bars indicate SEM.

Article Snippet: The primary antibodies were: anti-phospho p38 mitogen-activated protein kinase (p-p38 MAPK) alpha Thr180/Tyr182 antibody (Thermo Fisher Scientific; 36-8500, Rabbit, 1:500), anti-p38 MAPK antibody (Cell Signaling Technology; 9212, Rabbit, 1:1000), anti-dual-specificity phosphatases 1 (DUSP1) antibody (Cell Signaling Technology; 48625, Rabbit, 1:1000), anti-dual-specificity phosphatases 4 (DUSP4) antibody (Cell Signaling Technology; 5149, Rabbit, 1:1000), anti-dual-specificity phosphatases 12 (DUSP12) antibody (Proteintech, Wuhan, China; 67101-1-Ig, Mouse, 1:1000), anti-Poly/Mono-ADP ribose antibody (Cell Signaling Technology; 83732, Rabbit, 1:1000), anti-poly(ADP-ribose) polymerase family, member 1 (PARP1) antibody (Proteintech; 66520-1-Ig, Mouse, 1:1000), anti-mitogen-activated protein kinase 3 (MAP2K3) antibody (Proteintech; 80137-1-RR, Rabbit, 1:1000), anti-CPT1A antibody (Abcam; ab234111, Rabbit, 1:1000), anti-CPT1B antibody (Proteintech; 22170-1-AP, Rabbit, 1:1000), and anti-glucokinase (GCK) antibody (Proteintech; 19666-1-AP, Rabbit, 1:1000).

Techniques: Inhibition, Expressing, Phospho-proteomics, Control

CPT1 inhibition suppresses p38 MAPK phosphorylation mediated by PARP1 regulated ADP-ribosylation. (A) The relative abundance of metabolites extracted from octuplicate samples of control and ETX-treated heart ( n = 8). (B) Volcano plot for the control and ETX-treated hearts and the top 5 metabolites in up or down-regulated metabolites are shown ( n = 8). (C) The effect of ETX on poly(ADP-ribose) (PAR) protein in P7 cardiomyocytes determined by immunoblotting ( n = 6, ∗ P < 0.05 vs. Control). (D) Parp1 , poly(ADP-ribose) polymerase 2 ( Parp2 ), poly(ADP-ribose) polymerase 3 ( Parp3 ), poly(ADP-ribose) polymerase 4 ( Parp4 ), poly(ADP-ribose) polymerase 6 ( Parp6 ), poly(ADP-ribose) polymerase 8 ( Parp8 ), poly(ADP-ribose) polymerase 11 ( Parp11 ), poly(ADP-ribose) polymerase 12 ( Parp12 ), poly(ADP-ribose) polymerase 13 ( Parp13 ), poly(ADP-ribose) polymerase 16 ( Parp16 ), ADP-ribosyltransferase 1 ( Art1 ), ADP-ribosyltransferase 3 ( Art3 ), ADP-ribosyltransferase 4 ( Art4 ), mono-ADP ribosylhydrolase 1 ( Macrod1 ), mono-ADP ribosylhydrolase 2 ( Macrod2 ), ADP-ribosylserine hydrolase ( Adprs ), and poly(ADP-ribose) glycohydrolase ( Parg ) gene expression with ETX or without (Control) ETX treatment in P7 cardiomyocytes ( n = 6, ∗ P < 0.05 vs. Control). (E) The effect of ETX treatment and PARP1 overexpression (PARP1 OE) on PARP1 and PAR expression in P7 cardiomyocytes ( n = 6, ∗ P < 0.05, NS=Not significant). (F) The effect of ETX treatment and/or PARP1 OE on PARP1 expression and p38 MPAK phosphorylation in P7 cardiomyocytes ( n = 3, ∗ P < 0.05, NS=Not significant). (G, H) The effect of ETX treatment and PARP1 OE on the ratio of Ki67 + (red, white arrow) or PH3 + (red, white arrow) in P7 cardiomyocytes ( n = 6, ∗ P < 0.05, NS=Not significant; Scale bars = 50 μm). Error bars indicate SEM.

Journal: Acta Pharmaceutica Sinica. B

Article Title: Reversing metabolic reprogramming by CPT1 inhibition with etomoxir promotes cardiomyocyte proliferation and heart regeneration via DUSP1 ADP-ribosylation-mediated p38 MAPK phosphorylation

doi: 10.1016/j.apsb.2024.11.001

Figure Lengend Snippet: CPT1 inhibition suppresses p38 MAPK phosphorylation mediated by PARP1 regulated ADP-ribosylation. (A) The relative abundance of metabolites extracted from octuplicate samples of control and ETX-treated heart ( n = 8). (B) Volcano plot for the control and ETX-treated hearts and the top 5 metabolites in up or down-regulated metabolites are shown ( n = 8). (C) The effect of ETX on poly(ADP-ribose) (PAR) protein in P7 cardiomyocytes determined by immunoblotting ( n = 6, ∗ P < 0.05 vs. Control). (D) Parp1 , poly(ADP-ribose) polymerase 2 ( Parp2 ), poly(ADP-ribose) polymerase 3 ( Parp3 ), poly(ADP-ribose) polymerase 4 ( Parp4 ), poly(ADP-ribose) polymerase 6 ( Parp6 ), poly(ADP-ribose) polymerase 8 ( Parp8 ), poly(ADP-ribose) polymerase 11 ( Parp11 ), poly(ADP-ribose) polymerase 12 ( Parp12 ), poly(ADP-ribose) polymerase 13 ( Parp13 ), poly(ADP-ribose) polymerase 16 ( Parp16 ), ADP-ribosyltransferase 1 ( Art1 ), ADP-ribosyltransferase 3 ( Art3 ), ADP-ribosyltransferase 4 ( Art4 ), mono-ADP ribosylhydrolase 1 ( Macrod1 ), mono-ADP ribosylhydrolase 2 ( Macrod2 ), ADP-ribosylserine hydrolase ( Adprs ), and poly(ADP-ribose) glycohydrolase ( Parg ) gene expression with ETX or without (Control) ETX treatment in P7 cardiomyocytes ( n = 6, ∗ P < 0.05 vs. Control). (E) The effect of ETX treatment and PARP1 overexpression (PARP1 OE) on PARP1 and PAR expression in P7 cardiomyocytes ( n = 6, ∗ P < 0.05, NS=Not significant). (F) The effect of ETX treatment and/or PARP1 OE on PARP1 expression and p38 MPAK phosphorylation in P7 cardiomyocytes ( n = 3, ∗ P < 0.05, NS=Not significant). (G, H) The effect of ETX treatment and PARP1 OE on the ratio of Ki67 + (red, white arrow) or PH3 + (red, white arrow) in P7 cardiomyocytes ( n = 6, ∗ P < 0.05, NS=Not significant; Scale bars = 50 μm). Error bars indicate SEM.

Article Snippet: The primary antibodies were: anti-phospho p38 mitogen-activated protein kinase (p-p38 MAPK) alpha Thr180/Tyr182 antibody (Thermo Fisher Scientific; 36-8500, Rabbit, 1:500), anti-p38 MAPK antibody (Cell Signaling Technology; 9212, Rabbit, 1:1000), anti-dual-specificity phosphatases 1 (DUSP1) antibody (Cell Signaling Technology; 48625, Rabbit, 1:1000), anti-dual-specificity phosphatases 4 (DUSP4) antibody (Cell Signaling Technology; 5149, Rabbit, 1:1000), anti-dual-specificity phosphatases 12 (DUSP12) antibody (Proteintech, Wuhan, China; 67101-1-Ig, Mouse, 1:1000), anti-Poly/Mono-ADP ribose antibody (Cell Signaling Technology; 83732, Rabbit, 1:1000), anti-poly(ADP-ribose) polymerase family, member 1 (PARP1) antibody (Proteintech; 66520-1-Ig, Mouse, 1:1000), anti-mitogen-activated protein kinase 3 (MAP2K3) antibody (Proteintech; 80137-1-RR, Rabbit, 1:1000), anti-CPT1A antibody (Abcam; ab234111, Rabbit, 1:1000), anti-CPT1B antibody (Proteintech; 22170-1-AP, Rabbit, 1:1000), and anti-glucokinase (GCK) antibody (Proteintech; 19666-1-AP, Rabbit, 1:1000).

Techniques: Inhibition, Phospho-proteomics, Control, Western Blot, Gene Expression, Over Expression, Expressing

CPT1 inhibition suppresses p38 MAPK phosphorylation mediated by ADP-ribosylation of DUSP1. (A) The effect of ETX treatment on the interaction between DUSP1 and PARP1 in P7 cardiomyocytes ( n = 4, ∗ P < 0.05 vs. Control). (B) The effect of ETX treatment on the on ADP-ribosylation of DUSP1 in P7 cardiomyocytes ( n = 4, ∗ P < 0.05 vs. Control). (C) The effect of ETX treatment and/or PARP1 OE on ADP-ribosylation of DUSP1 and the interaction between DUSP1 and p38 MAPK in P7 cardiomyocytes ( n = 3, ∗ P < 0.05, NS=Not significant). (D) LVEF, LVFS, LVIDd and LVIDs at 4 weeks after MI were measured in MADM mice after treatment with ETX and/or PARP1 AAV9 (PARP1 overexpression, PARP1 OE) ( n = 5, ∗ P < 0.05, NS=Not significant). (E) The infarcted area of the heart in mice 4 weeks after MI treated with control (Vector), ETX, and/or PARP1 OE ( n = 5, ∗ P < 0.05, NS=Not significant; Scale bars = 1 mm). (F) Representative immunofluorescence and quantification of single-labeled (red or green, white arrow) cardiomyocytes in MADM mice after MI for 4 weeks treated with control (Vector), ETX, and/or PARP1 OE ( n = 5, ∗ P < 0.05, NS=Not significant; Scale bars = 20 μm). (G) The effect of ETX and PARP1 OE on ADP-ribosylation of DUSP1 and the interaction between DUSP1 and p38 MAPK in mice hearts ( n = 3, ∗ P < 0.05, NS=Not significant). Error bars indicate SEM.

Journal: Acta Pharmaceutica Sinica. B

Article Title: Reversing metabolic reprogramming by CPT1 inhibition with etomoxir promotes cardiomyocyte proliferation and heart regeneration via DUSP1 ADP-ribosylation-mediated p38 MAPK phosphorylation

doi: 10.1016/j.apsb.2024.11.001

Figure Lengend Snippet: CPT1 inhibition suppresses p38 MAPK phosphorylation mediated by ADP-ribosylation of DUSP1. (A) The effect of ETX treatment on the interaction between DUSP1 and PARP1 in P7 cardiomyocytes ( n = 4, ∗ P < 0.05 vs. Control). (B) The effect of ETX treatment on the on ADP-ribosylation of DUSP1 in P7 cardiomyocytes ( n = 4, ∗ P < 0.05 vs. Control). (C) The effect of ETX treatment and/or PARP1 OE on ADP-ribosylation of DUSP1 and the interaction between DUSP1 and p38 MAPK in P7 cardiomyocytes ( n = 3, ∗ P < 0.05, NS=Not significant). (D) LVEF, LVFS, LVIDd and LVIDs at 4 weeks after MI were measured in MADM mice after treatment with ETX and/or PARP1 AAV9 (PARP1 overexpression, PARP1 OE) ( n = 5, ∗ P < 0.05, NS=Not significant). (E) The infarcted area of the heart in mice 4 weeks after MI treated with control (Vector), ETX, and/or PARP1 OE ( n = 5, ∗ P < 0.05, NS=Not significant; Scale bars = 1 mm). (F) Representative immunofluorescence and quantification of single-labeled (red or green, white arrow) cardiomyocytes in MADM mice after MI for 4 weeks treated with control (Vector), ETX, and/or PARP1 OE ( n = 5, ∗ P < 0.05, NS=Not significant; Scale bars = 20 μm). (G) The effect of ETX and PARP1 OE on ADP-ribosylation of DUSP1 and the interaction between DUSP1 and p38 MAPK in mice hearts ( n = 3, ∗ P < 0.05, NS=Not significant). Error bars indicate SEM.

Article Snippet: The primary antibodies were: anti-phospho p38 mitogen-activated protein kinase (p-p38 MAPK) alpha Thr180/Tyr182 antibody (Thermo Fisher Scientific; 36-8500, Rabbit, 1:500), anti-p38 MAPK antibody (Cell Signaling Technology; 9212, Rabbit, 1:1000), anti-dual-specificity phosphatases 1 (DUSP1) antibody (Cell Signaling Technology; 48625, Rabbit, 1:1000), anti-dual-specificity phosphatases 4 (DUSP4) antibody (Cell Signaling Technology; 5149, Rabbit, 1:1000), anti-dual-specificity phosphatases 12 (DUSP12) antibody (Proteintech, Wuhan, China; 67101-1-Ig, Mouse, 1:1000), anti-Poly/Mono-ADP ribose antibody (Cell Signaling Technology; 83732, Rabbit, 1:1000), anti-poly(ADP-ribose) polymerase family, member 1 (PARP1) antibody (Proteintech; 66520-1-Ig, Mouse, 1:1000), anti-mitogen-activated protein kinase 3 (MAP2K3) antibody (Proteintech; 80137-1-RR, Rabbit, 1:1000), anti-CPT1A antibody (Abcam; ab234111, Rabbit, 1:1000), anti-CPT1B antibody (Proteintech; 22170-1-AP, Rabbit, 1:1000), and anti-glucokinase (GCK) antibody (Proteintech; 19666-1-AP, Rabbit, 1:1000).

Techniques: Inhibition, Phospho-proteomics, Control, Over Expression, Plasmid Preparation, Immunofluorescence, Labeling

Schematic representation of the proposed effect of reversing metabolic reprogramming by CPT1 inhibition on cardiomyocyte proliferation and heart regeneration. Inhibition of FAO by CPT1 KO or Etomoxir decreases PARP1 expression, negatively regulates ADP-ribosylation modification of DUSP1 and dephosphorylates p38 MAPK which increases cell cycle gene expression and promotes cardiomyocyte proliferation. The suppression of postnatal cardiomyocyte cell-cycle arrest increases cardiac regeneration post myocardial infraction injury.

Journal: Acta Pharmaceutica Sinica. B

Article Title: Reversing metabolic reprogramming by CPT1 inhibition with etomoxir promotes cardiomyocyte proliferation and heart regeneration via DUSP1 ADP-ribosylation-mediated p38 MAPK phosphorylation

doi: 10.1016/j.apsb.2024.11.001

Figure Lengend Snippet: Schematic representation of the proposed effect of reversing metabolic reprogramming by CPT1 inhibition on cardiomyocyte proliferation and heart regeneration. Inhibition of FAO by CPT1 KO or Etomoxir decreases PARP1 expression, negatively regulates ADP-ribosylation modification of DUSP1 and dephosphorylates p38 MAPK which increases cell cycle gene expression and promotes cardiomyocyte proliferation. The suppression of postnatal cardiomyocyte cell-cycle arrest increases cardiac regeneration post myocardial infraction injury.

Article Snippet: The primary antibodies were: anti-phospho p38 mitogen-activated protein kinase (p-p38 MAPK) alpha Thr180/Tyr182 antibody (Thermo Fisher Scientific; 36-8500, Rabbit, 1:500), anti-p38 MAPK antibody (Cell Signaling Technology; 9212, Rabbit, 1:1000), anti-dual-specificity phosphatases 1 (DUSP1) antibody (Cell Signaling Technology; 48625, Rabbit, 1:1000), anti-dual-specificity phosphatases 4 (DUSP4) antibody (Cell Signaling Technology; 5149, Rabbit, 1:1000), anti-dual-specificity phosphatases 12 (DUSP12) antibody (Proteintech, Wuhan, China; 67101-1-Ig, Mouse, 1:1000), anti-Poly/Mono-ADP ribose antibody (Cell Signaling Technology; 83732, Rabbit, 1:1000), anti-poly(ADP-ribose) polymerase family, member 1 (PARP1) antibody (Proteintech; 66520-1-Ig, Mouse, 1:1000), anti-mitogen-activated protein kinase 3 (MAP2K3) antibody (Proteintech; 80137-1-RR, Rabbit, 1:1000), anti-CPT1A antibody (Abcam; ab234111, Rabbit, 1:1000), anti-CPT1B antibody (Proteintech; 22170-1-AP, Rabbit, 1:1000), and anti-glucokinase (GCK) antibody (Proteintech; 19666-1-AP, Rabbit, 1:1000).

Techniques: Inhibition, Expressing, Modification, Gene Expression

Fig. 11. Endogenous tau levels are decreased following treatment with the Syk inhibitor BAY 61–3606. Endogenous total tau (Tau5) and phosphorylated tau (PHF-1) levels are decreased in noninduced M1C cells following treatment with the Syk inhibitor BAY 61–3606. Data are presented as mean ± SD (N = 4). All data were normally distributed and were analyzed using Student’s t-test. *p < 0.05, **p < 0.01. Abbreviations: GAPDH, glyceraldehyde 3-phosphate dehy- drogenase; LC3, microtubule-associated protein 1 light chain 3; SD, standard deviation; Syk, splenic tyrosine kinase.

Journal: Neurobiology of disease

Article Title: Syk inhibitors reduce tau protein phosphorylation and oligomerization.

doi: 10.1016/j.nbd.2024.106656

Figure Lengend Snippet: Fig. 11. Endogenous tau levels are decreased following treatment with the Syk inhibitor BAY 61–3606. Endogenous total tau (Tau5) and phosphorylated tau (PHF-1) levels are decreased in noninduced M1C cells following treatment with the Syk inhibitor BAY 61–3606. Data are presented as mean ± SD (N = 4). All data were normally distributed and were analyzed using Student’s t-test. *p < 0.05, **p < 0.01. Abbreviations: GAPDH, glyceraldehyde 3-phosphate dehy- drogenase; LC3, microtubule-associated protein 1 light chain 3; SD, standard deviation; Syk, splenic tyrosine kinase.

Article Snippet: Antibodies against phosphorylated (p)p38MAPK (Thr180/Tyr182), p38MAPK, and microtubule-associated protein 1 light chain 3 (LC3) were obtained from Cell Signaling (#9211, #92112, and #3868; Danvers, MA, USA).

Techniques: Standard Deviation

Molecular Regulation of Fibrosis in Ex Vivo Cultured Hearts Representative pictures of the right ventricle and the right atrium of hearts cultured for 7 days under standard conditions (−), in the presence of SB431542 (SB, SB1, SB2) or transforming growth factor-beta (TGF-β) for (A) phospho-SMAD2/3 (pSMAD2/3) and phospho-p38 mitogen-activated protein kinase (pp38MAPK), (B) α-SMA and vimentin, (C) PECAM-1 and cTnI, (D) collagen I and collagen III, and (E) Alcian blue, periostin, versican B, and fibronectin. (F) Graphs depicting the percentage of hearts with α-SMA expression in the subepicardium of the ventricle and atrium after culture for 7 days under standard conditions (−), in the presence of SB431542 (SB) or TGF-β. (G and H) Graphs depicting the relative α-SMA-positive area (G) and the relative vimentin-positive area (H) in the ventricular or atrial myocardium of hearts cultured for 7 days under standard conditions (−), in the presence of SB431542 (SB) or TGF-β. DAPI was used as counterstain in all fluorescent pictures. Data are presented as means ± SEM. To evaluate significant differences, 1-way analysis of variance followed by Tukey’s multiple comparisons test was performed for graphs (G and H). Kruskal-Wallis test followed by Dunn’s multiple comparisons test was performed for graph (G [atrium]). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Scale bar: 20 μm. Abbreviations as in <xref ref-type=Figure 1 . " width="100%" height="100%">

Journal: JACC: Basic to Translational Science

Article Title: A New Ex Vivo Model to Study Cardiac Fibrosis in Whole Mouse Hearts

doi: 10.1016/j.jacbts.2024.04.007

Figure Lengend Snippet: Molecular Regulation of Fibrosis in Ex Vivo Cultured Hearts Representative pictures of the right ventricle and the right atrium of hearts cultured for 7 days under standard conditions (−), in the presence of SB431542 (SB, SB1, SB2) or transforming growth factor-beta (TGF-β) for (A) phospho-SMAD2/3 (pSMAD2/3) and phospho-p38 mitogen-activated protein kinase (pp38MAPK), (B) α-SMA and vimentin, (C) PECAM-1 and cTnI, (D) collagen I and collagen III, and (E) Alcian blue, periostin, versican B, and fibronectin. (F) Graphs depicting the percentage of hearts with α-SMA expression in the subepicardium of the ventricle and atrium after culture for 7 days under standard conditions (−), in the presence of SB431542 (SB) or TGF-β. (G and H) Graphs depicting the relative α-SMA-positive area (G) and the relative vimentin-positive area (H) in the ventricular or atrial myocardium of hearts cultured for 7 days under standard conditions (−), in the presence of SB431542 (SB) or TGF-β. DAPI was used as counterstain in all fluorescent pictures. Data are presented as means ± SEM. To evaluate significant differences, 1-way analysis of variance followed by Tukey’s multiple comparisons test was performed for graphs (G and H). Kruskal-Wallis test followed by Dunn’s multiple comparisons test was performed for graph (G [atrium]). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Scale bar: 20 μm. Abbreviations as in Figure 1 .

Article Snippet: After blocking with 1% BSA in 0.1% Tween-PBS, sections were incubated overnight with the primary antibodies directed against platelet endothelial cell adhesion molecule (PECAM-1) (R&D, AF3628; 1:1,000), proliferating cell nuclear antigen (PCNA) (Sigma, P8825; 1:1,000), smooth muscle actin-alpha (α-SMA, Sigma; A2547; 1:20,000), collagen I (SouthernBiotech, 1310-01; 1:200), GFP (Abcam, ab13970, 1:1,000), collagen III (SouthernBiotech, 1330-01; 1:200), phospho-p38 mitogen-activated protein kinase (pp38MAPK) (Cell Signaling, #4631; 1:100), periostin (Santa Cruz, SC398631; 1:100), versican B (Chemicon, AB1033; 1:100), fibronectin (Sigma, F3648; 1:200), cardiac troponin-I (cTnI) (HyTest, 4T21/2; 1:1,000), matrix metalloproteinase 2 (MMP2) (Invitrogen, #436000; 1:200), vimentin (Cell Signaling, #5741; 1:200), mannose receptor (CD206, Abcam; ab64693; 1:300), F4/80 (Invitrogen, 14-4801-82; 1:100), MAC3 (CD107b, BD Biosciences, #550292; 1:200), CD68 (Santa Cruz, SC20060; 1:100), and sarcomeric myofilaments (MF20, Developmental Studies Hybridoma Bank, University of Iowa) followed by incubation with Alexa-conjugated secondary antibodies (Molecular Probes).

Techniques: Ex Vivo, Cell Culture, Expressing