phospho-mkp1/dusp1 (ser359, #2857) antibody  (Cell Signaling Technology Inc)

Journal: mBio

Article Title: Variations in candidalysin amino acid sequence influence toxicity and host responses

doi: 10.1128/mbio.03351-23

Figure Lengend Snippet: Candidalysin variants induce differential activation of MAPK signaling and cytokine secretion. Western blot analysis of TR146 epithelial cells treated with 15 µM of each candidalysin variant from C. albicans , C. dubliniensis, and C. tropicalis for ( A ) 30 min and ( B ) 2 h. Protein lysates (10 µg total protein) were probed with antibodies specific for MKK3, MKK6, and p-MKK3/6 ( A ), and p-EGFR, c-Fos, p-MKP1 ( B ). α-actin was used as a loading control. One representative blot presented from n = 3 biological repeats. Quantification of G-CSF, GM-CSF, IL-1α, IL-1β, and IL-6 secreted from TR146 epithelial cells treated with 15 µM of each candidalysin variant from ( C ) C. albicans , and ( D ) C. dubliniensis and C. tropicalis for 24 h. Data are the mean + SD of n = 3 biological repeats ( C. albicans candidalysin variant A vs variant B; no significant difference for all cytokines tested). Statistical analysis was applied relative to vehicle-treated cells using a one-way ANOVA with a post hoc Bonferroni multiple comparison test; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Primary antibodies specific for MKK3 (#8535), MKK6 (#8550), p-MKK3/6 (MKK3-Ser189 and MKK6-Ser207; #12280), p-EGFR Y1068 (#3777S), c-Fos (#2250S), and p-MKP1 (#2857S) were purchased from Cell Signaling Technology. α-Actin (#MAB1501) was purchased from Millipore.

Techniques: Activation Assay, Western Blot, Variant Assay, Control, Comparison

Journal: PLOS Pathogens

Article Title: Candida albicans translocation through the intestinal epithelial barrier is promoted by fungal zinc acquisition and limited by NFκB-mediated barrier protection

doi: 10.1371/journal.ppat.1012031

Figure Lengend Snippet: Detection antibodies for immune signaling proteins.

Article Snippet: p-DUSP1/MKP1 (S359) , Rabbit , 1:1,000 , Cell Signaling , 2857.

Techniques:

Journal: Journal of Cancer

Article Title: A mechanism study of DUSP1 in inhibiting malignant progression of endometrial carcinoma by regulating ERK/AP-1 axis and dephosphorylation of EPHA2

doi: 10.7150/jca.81069

Figure Lengend Snippet: Sequences of primers and siRNAs.

Article Snippet: The antibodies against DUSP1 (#2857S), ERK1/2 (#4695S), p-ERK1/2 (4370S), EPHA2 (6997S), GAPDH (#2118S) were all purchased from CST (Cell Signaling Technology, Danvers, USA) and antibodies against JNK (#51151-1-AP) and p-JNK (#80024-1-RR) were purchased from ProteinTech (Rosemont, USA).

Techniques: Sequencing

Journal: Journal of Cancer

Article Title: A mechanism study of DUSP1 in inhibiting malignant progression of endometrial carcinoma by regulating ERK/AP-1 axis and dephosphorylation of EPHA2

doi: 10.7150/jca.81069

Figure Lengend Snippet: Co-expression analysis of DUSP1 in EC.

Article Snippet: The antibodies against DUSP1 (#2857S), ERK1/2 (#4695S), p-ERK1/2 (4370S), EPHA2 (6997S), GAPDH (#2118S) were all purchased from CST (Cell Signaling Technology, Danvers, USA) and antibodies against JNK (#51151-1-AP) and p-JNK (#80024-1-RR) were purchased from ProteinTech (Rosemont, USA).

Techniques:

Journal: Journal of Cancer

Article Title: A mechanism study of DUSP1 in inhibiting malignant progression of endometrial carcinoma by regulating ERK/AP-1 axis and dephosphorylation of EPHA2

doi: 10.7150/jca.81069

Figure Lengend Snippet: Correlation analysis of DUSP1 in endometrial carcinoma. Genes with the highest correlation with DUSP1 in EC were selected in this study. Individual Pearson coefficient and P values were marked on the figures.

Article Snippet: The antibodies against DUSP1 (#2857S), ERK1/2 (#4695S), p-ERK1/2 (4370S), EPHA2 (6997S), GAPDH (#2118S) were all purchased from CST (Cell Signaling Technology, Danvers, USA) and antibodies against JNK (#51151-1-AP) and p-JNK (#80024-1-RR) were purchased from ProteinTech (Rosemont, USA).

Techniques:

Journal: Journal of Cancer

Article Title: A mechanism study of DUSP1 in inhibiting malignant progression of endometrial carcinoma by regulating ERK/AP-1 axis and dephosphorylation of EPHA2

doi: 10.7150/jca.81069

Figure Lengend Snippet: Modulation of DUSP1 affects the expression of AP-1 genes. (A) mRNA and protein expression level after transfection with different DUSP1 siRNAs. (B) mRNA and protein expression level after transfection with plasmid PC-h-DUSP1. (C) Expression of selected genes in Ishikawa cells after transfection of DUSP1. (D) Overexpression of DUSP1 dephosphorylate MAPK pathway. 1: negative control group (NC), 2: knockdown of DUSP1 in Ishikawa 3: overexpression of DUSP1 in Ishikawa; Relative expression: the relative expression calculated based on ΔCT method; Si1 # : DUSP1 SiRNA1; Si2 # : DUSP1 SiRNA2; Si3 # : DUSP1 SiRNA3; PC-h-DUSP1: plasmid PC-h-DUSP1 for overexpression; NC: negative control; *: p<0.05; **: p<0.01; ***: p<0.001; ****: p<0.0001.

Article Snippet: The antibodies against DUSP1 (#2857S), ERK1/2 (#4695S), p-ERK1/2 (4370S), EPHA2 (6997S), GAPDH (#2118S) were all purchased from CST (Cell Signaling Technology, Danvers, USA) and antibodies against JNK (#51151-1-AP) and p-JNK (#80024-1-RR) were purchased from ProteinTech (Rosemont, USA).

Techniques: Expressing, Transfection, Plasmid Preparation, Over Expression, Negative Control, Knockdown

Journal: Journal of Cancer

Article Title: A mechanism study of DUSP1 in inhibiting malignant progression of endometrial carcinoma by regulating ERK/AP-1 axis and dephosphorylation of EPHA2

doi: 10.7150/jca.81069

Figure Lengend Snippet: AP-1 inhibitor could affect the expression of DUSP1. (A) Prediction of transcription factor binding sites on the promoter of DUSP1. (B) mRNA expression of DUSP1 after using different concentrations of AP-1 inhibitor. (C) protein expression of DUSP1 after using different concentrations of AP-1 inhibitor. Relative expression: the relative expression calculated based on ΔCT method; ***: p<0.001.

Article Snippet: The antibodies against DUSP1 (#2857S), ERK1/2 (#4695S), p-ERK1/2 (4370S), EPHA2 (6997S), GAPDH (#2118S) were all purchased from CST (Cell Signaling Technology, Danvers, USA) and antibodies against JNK (#51151-1-AP) and p-JNK (#80024-1-RR) were purchased from ProteinTech (Rosemont, USA).

Techniques: Expressing, Binding Assay

Journal: Journal of Cancer

Article Title: A mechanism study of DUSP1 in inhibiting malignant progression of endometrial carcinoma by regulating ERK/AP-1 axis and dephosphorylation of EPHA2

doi: 10.7150/jca.81069

Figure Lengend Snippet: Transfection of DUSP1 altered the cell apoptosis and viability of EC cell lines. (A) Cell apoptosis of different EC cell lines after DUSP1 transfection. (B) Statistics of apoptosis after transfection. (C) Proliferation ability of three kinds of EC cells transfected with DUSP1. Si1 # : DUSP1 SiRNA1; PC-h-DUSP1: plasmid PC-h-DUSP1 for overexpression; NC: negative control; **: p<0.01; ***: p<0.001.

Article Snippet: The antibodies against DUSP1 (#2857S), ERK1/2 (#4695S), p-ERK1/2 (4370S), EPHA2 (6997S), GAPDH (#2118S) were all purchased from CST (Cell Signaling Technology, Danvers, USA) and antibodies against JNK (#51151-1-AP) and p-JNK (#80024-1-RR) were purchased from ProteinTech (Rosemont, USA).

Techniques: Transfection, Plasmid Preparation, Over Expression, Negative Control

Journal: Journal of Cancer

Article Title: A mechanism study of DUSP1 in inhibiting malignant progression of endometrial carcinoma by regulating ERK/AP-1 axis and dephosphorylation of EPHA2

doi: 10.7150/jca.81069

Figure Lengend Snippet: Regulation of DUSP1 changed the invasion ability of endometrial carcinoma. (A) Representative pictures of transwell invasion assay after DUSP1 transfection. a1~a3 : control group, knockdown of DUSP1 and overexpression of DUSP1 in Ishikawa cell line; b1~b3 : control group, knockdown of DUSP1 and overexpression of DUSP1 in HEC-1B cell line; c1~c3 : control group, knockdown of DUSP1 and overexpression of DUSP1 in HEC-50B cell line. The scale bar represents 100 μm. (B) The number of transmembrane cells.

Article Snippet: The antibodies against DUSP1 (#2857S), ERK1/2 (#4695S), p-ERK1/2 (4370S), EPHA2 (6997S), GAPDH (#2118S) were all purchased from CST (Cell Signaling Technology, Danvers, USA) and antibodies against JNK (#51151-1-AP) and p-JNK (#80024-1-RR) were purchased from ProteinTech (Rosemont, USA).

Techniques: Transwell Invasion Assay, Transfection, Control, Knockdown, Over Expression

Journal: Journal of Cancer

Article Title: A mechanism study of DUSP1 in inhibiting malignant progression of endometrial carcinoma by regulating ERK/AP-1 axis and dephosphorylation of EPHA2

doi: 10.7150/jca.81069

Figure Lengend Snippet: Phosphorylation omics analysis after DUSP1 overexpression. (A) Distribution of phosphorylation amino acids of DUSP1. S: serine; T: threonine; Y: tyrosine. (B) Statistics and histogram of quantitative difference of phosphorylated peptides. (C) Volcano plot. (D) Main phosphorylated or dephosphorylated kinases after DUSP1 overexpression. NC: negative control.

Article Snippet: The antibodies against DUSP1 (#2857S), ERK1/2 (#4695S), p-ERK1/2 (4370S), EPHA2 (6997S), GAPDH (#2118S) were all purchased from CST (Cell Signaling Technology, Danvers, USA) and antibodies against JNK (#51151-1-AP) and p-JNK (#80024-1-RR) were purchased from ProteinTech (Rosemont, USA).

Techniques: Phospho-proteomics, Over Expression, Negative Control

Journal: Journal of Cancer

Article Title: A mechanism study of DUSP1 in inhibiting malignant progression of endometrial carcinoma by regulating ERK/AP-1 axis and dephosphorylation of EPHA2

doi: 10.7150/jca.81069

Figure Lengend Snippet: Effect of EPHA2 on tumor growth in endometrial carcinoma. (A) Co-IP detection of DUSP1 and EPHA2 (B) mRNA expression of EPHA2 after DUSP1 transfection. Relative expression: the relative expression calculated based on ΔCT method; **: p<0.01; ***: p<0.001. (C) protein expression of EPHA2 after DUSP1 transfection. 1: negative control group; 2: knockdown of DUSP1 in Ishikawa cell line; 3: overexpression of DUSP1 in Ishikawa cell line. (D) Representative pictures of EPHA2 expression detected by IHC. (E) Western blotting results of EPHA2 in EC tissues. G1 I : Grade 1, stage I, endometrioid; G2 II : Grade 2, stage II, endometrioid; G3 III : Grade 3, stage III, endometrioid. (F) growth curves after using EPHA2 inhibitor on Ishikawa.

Article Snippet: The antibodies against DUSP1 (#2857S), ERK1/2 (#4695S), p-ERK1/2 (4370S), EPHA2 (6997S), GAPDH (#2118S) were all purchased from CST (Cell Signaling Technology, Danvers, USA) and antibodies against JNK (#51151-1-AP) and p-JNK (#80024-1-RR) were purchased from ProteinTech (Rosemont, USA).

Techniques: Co-Immunoprecipitation Assay, Expressing, Transfection, Negative Control, Knockdown, Over Expression, Western Blot

Journal: eLife

Article Title: The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells

doi: 10.7554/eLife.75490

Figure Lengend Snippet: ( A ) Representative western blots of oral epithelial cells (TR146) probed for c-Fos expression and phosphorylation of MKP1 after 2 hr treatment with WT and variant CL. ( B ) Quantification of c-Fos and MKP1 phosphorylation was used to assess danger response signaling. Data were normalized to values obtained in control conditions. Actin was used as a loading control. N=4, and bars are the S.D. Figure 7—source data 1. Uncropped western blot of TR146 cell lysates treated with WT and variant CL for two hours probed with antibodies for (A) c-Fos, actin and (B) pMKP1.

Article Snippet: Blots were probed with primary antibodies for c-Fos (1:500, Cell Signaling Technology, 2250 S) and phospho-MKP1 (1:1000, Cell Signaling Technology, 2857 S) overnight.

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

Journal: eLife

Article Title: The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells

doi: 10.7554/eLife.75490

Figure Lengend Snippet:

Article Snippet: Blots were probed with primary antibodies for c-Fos (1:500, Cell Signaling Technology, 2250 S) and phospho-MKP1 (1:1000, Cell Signaling Technology, 2857 S) overnight.

Techniques: Software, Recombinant

Journal: eLife

Article Title: The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells

doi: 10.7554/elife.75490

Figure Lengend Snippet: Figure 7. The polymerization-defective G4W variant does not cause danger response signaling in epithelial cells. (A) Representative western blots of oral epithelial cells (TR146) probed for c-Fos expression and phosphorylation of MKP1 after 2 hr treatment with WT and variant CL. (B) Quantification of c-Fos and MKP1 phosphorylation was used to assess danger response signaling. Data were normalized to values obtained in control conditions. Actin was used as a loading control. N=4, and bars are the S.D.

Article Snippet: DOI: https://doi.org/10.7554/eLife.75490 14 of 21 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Cell line (Homo sapiens) Buccal squamous cell carcinoma ECACC TR146 ECACC 10032305 Antibody Goat- anti mouse IRDye 680 (Goat polyclonal) LI- COR 925–68070 1:10,000 Antibody Phospho- MKP1 (Rabbit monoclonal) Cell Signaling Technology 2857 S 1:1,000 Antibody c- Fos (Rabbit monoclonal) Cell Signaling Technology 2250 S 1:500 Antibody Goat- anti rabbit IRDye 800 (Goat polyclonal) LI- COR 926–32211 1:10,000 Antibody β-actin (mouse monoclonal) ABCAM ab6276 1:1000 Peptide, recombinant protein Wild- Type Candidalysin Peptide 2.0 CL Peptide, recombinant protein I9A Candidalysin Peptide 2.0 I9A Peptide, recombinant protein G4W Candidalysin Peptide 2.0 G4W Chemical compound, drug POPC Avanti Polar Lipids 850457 C Chemical compound, drug DOPC Avanti Polar Lipids 850375 P Chemical compound, drug Calcein MP 02190167- CF Software, algorithm Discover MP Refeyn version 2.2.0 Software, algorithm Acquire MP Refeyn version 2.2.0 Software, algorithm SEDFIT NIH Software, algorithm Image Studio LI- COR Chemical compound, drug n- Dodecyl- beta- DMaltoside Millipore Sigma D4641 DBM Software, algorithm Igor Pro 7.08 Wavemetrics Software, algorithm Igor Pro 6.38 Wavemetrics Software, algorithm Asylum AFM Software Oxford instruments Version 16 Other AFM tips Olympus See AFM Materials and methods Section Continued

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