ask1 d11c9  (Cell Signaling Technology Inc)

Journal: International Journal of Molecular Medicine

Article Title: Kirenol relieves rheumatoid arthritis by targeting the TWEAK/Fn14 pathway

doi: 10.3892/ijmm.2025.5586

Figure Lengend Snippet: Kir attenuates the levels of TWEAK/Fn14 pathway-associated proteins in the synovial tissues of CIA rats. Representative western blots and quantification of the (A) TWEAK, (B) Fn14, (C) TRAF2, (D) ASK1 and (E) NIK protein levels and the phosphorylation of (F) IRE1, (G) IKKα and (H) p100. β-actin was used as the loading control. All the data are expressed as the mean ± SD (n=5). Kruskal-Wallis followed by Dunn's post-test was performed in D, one-way ANOVA followed by Games-Howell in C and one-way ANOVA followed by Tukey test was performed in A, B and E-H. ** P<0.01 vs. the control group, # P<0.05 or ## P<0.01 vs. the CIA group. Fn14, fibroblast growth factor-inducible immediate-early response protein 14; Kir, kirenol; CIA, collagen-induced arthritis; TWEAK, tumor necrosis factor-like weak inducer of apoptosis; NIK, nuclear factor-κB-inducing kinase; IRE1, inositol-requiring enzyme 1; IKKα, inhibitor of κB kinase α.

Article Snippet: ASK1 (D11C9) , Cell Signaling Technology, Inc. , 8662 , WB/1:800.

Techniques: Western Blot, Phospho-proteomics, Control

Journal: Biology of Reproduction

Article Title: Oxidative stress-induced TGF-beta/TAB1-mediated p38MAPK activation in human amnion epithelial cells †

doi: 10.1093/biolre/ioy135

Figure Lengend Snippet: Antibodies used for the western blot.

Article Snippet: ASK1 , D11C9 , Cell Signaling , 1:750 , Western Blot.

Techniques: Western Blot, Immunofluorescence, Immunoprecipitation

Journal: Biology of Reproduction

Article Title: Oxidative stress-induced TGF-beta/TAB1-mediated p38MAPK activation in human amnion epithelial cells †

doi: 10.1093/biolre/ioy135

Figure Lengend Snippet: List of siRNA target sequences.

Article Snippet: The following antihuman antibodies were used for western blot: ASK1 (1:750, Cell Signaling, Danvers, MA), TRX (1:1000, Abcam, Cambridge, United Kingdom), TAB1 (1:1000, R&D, Minneapolis, MN), P-TAB1 (1:800, Milipore, Thr431, Burlington, MA), P-p38MAPK (1:300, Cell Signaling, T180/Y182), p38MAPK (1:1000, Cell Signaling) (Table ). table ft1 table-wrap mode="anchored" t5 Table 3. caption a7 Protein Name Catalog Number Company Dilution Method ASK1 D11C9 Cell Signaling 1:750 Western Blot TRX Ab16965 Abcam 1:1000 Western Blot TAB1 AF3578 R&D 1:1000 Western Blot P-TAB1(Thr431) 06–1334 Milipore 1:800 Western Blot P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Western Blot p38 MAPK 9212S Cell Signaling 1:1000 Western Blot Β-Actin A5441 Sigma-Aldrich 1:20,000 Western Blot Cytokeratin-18 Ab668 Abcam 1:300 Immunofluorescence ASK1 Ab45178 Abcam 1:250 Immunofluorescence TRX Ab16965 Abcam 1:500 Immunofluorescence P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Immunofluorescence TAK1 Sc-7967 Santa Cruz 1:300 Immunofluorescence TAB1 AF3578 R&D 1:300 Immunofluorescence ASK1 D11C9 Cell Signaling 1:100 Immunoprecipitation TRX Ab16965 Abcam 1:100 Immunoprecipitation GSK3B mAb#9832 Cell Signaling 1:100 Immunoprecipitation Open in a separate window Antibodies used for the western blot.

Techniques: Sequencing

Journal: Biology of Reproduction

Article Title: Oxidative stress-induced TGF-beta/TAB1-mediated p38MAPK activation in human amnion epithelial cells †

doi: 10.1093/biolre/ioy135

Figure Lengend Snippet: List of qPCR assays primers for AEC experiments.

Article Snippet: The following antihuman antibodies were used for western blot: ASK1 (1:750, Cell Signaling, Danvers, MA), TRX (1:1000, Abcam, Cambridge, United Kingdom), TAB1 (1:1000, R&D, Minneapolis, MN), P-TAB1 (1:800, Milipore, Thr431, Burlington, MA), P-p38MAPK (1:300, Cell Signaling, T180/Y182), p38MAPK (1:1000, Cell Signaling) (Table ). table ft1 table-wrap mode="anchored" t5 Table 3. caption a7 Protein Name Catalog Number Company Dilution Method ASK1 D11C9 Cell Signaling 1:750 Western Blot TRX Ab16965 Abcam 1:1000 Western Blot TAB1 AF3578 R&D 1:1000 Western Blot P-TAB1(Thr431) 06–1334 Milipore 1:800 Western Blot P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Western Blot p38 MAPK 9212S Cell Signaling 1:1000 Western Blot Β-Actin A5441 Sigma-Aldrich 1:20,000 Western Blot Cytokeratin-18 Ab668 Abcam 1:300 Immunofluorescence ASK1 Ab45178 Abcam 1:250 Immunofluorescence TRX Ab16965 Abcam 1:500 Immunofluorescence P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Immunofluorescence TAK1 Sc-7967 Santa Cruz 1:300 Immunofluorescence TAB1 AF3578 R&D 1:300 Immunofluorescence ASK1 D11C9 Cell Signaling 1:100 Immunoprecipitation TRX Ab16965 Abcam 1:100 Immunoprecipitation GSK3B mAb#9832 Cell Signaling 1:100 Immunoprecipitation Open in a separate window Antibodies used for the western blot.

Techniques:

Journal: Biology of Reproduction

Article Title: Oxidative stress-induced TGF-beta/TAB1-mediated p38MAPK activation in human amnion epithelial cells †

doi: 10.1093/biolre/ioy135

Figure Lengend Snippet: Antibodies used for the western blot.

Article Snippet: The following antihuman antibodies were used for western blot: ASK1 (1:750, Cell Signaling, Danvers, MA), TRX (1:1000, Abcam, Cambridge, United Kingdom), TAB1 (1:1000, R&D, Minneapolis, MN), P-TAB1 (1:800, Milipore, Thr431, Burlington, MA), P-p38MAPK (1:300, Cell Signaling, T180/Y182), p38MAPK (1:1000, Cell Signaling) (Table ). table ft1 table-wrap mode="anchored" t5 Table 3. caption a7 Protein Name Catalog Number Company Dilution Method ASK1 D11C9 Cell Signaling 1:750 Western Blot TRX Ab16965 Abcam 1:1000 Western Blot TAB1 AF3578 R&D 1:1000 Western Blot P-TAB1(Thr431) 06–1334 Milipore 1:800 Western Blot P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Western Blot p38 MAPK 9212S Cell Signaling 1:1000 Western Blot Β-Actin A5441 Sigma-Aldrich 1:20,000 Western Blot Cytokeratin-18 Ab668 Abcam 1:300 Immunofluorescence ASK1 Ab45178 Abcam 1:250 Immunofluorescence TRX Ab16965 Abcam 1:500 Immunofluorescence P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Immunofluorescence TAK1 Sc-7967 Santa Cruz 1:300 Immunofluorescence TAB1 AF3578 R&D 1:300 Immunofluorescence ASK1 D11C9 Cell Signaling 1:100 Immunoprecipitation TRX Ab16965 Abcam 1:100 Immunoprecipitation GSK3B mAb#9832 Cell Signaling 1:100 Immunoprecipitation Open in a separate window Antibodies used for the western blot.

Techniques: Western Blot, Immunofluorescence, Immunoprecipitation

Journal: Biology of Reproduction

Article Title: Oxidative stress-induced TGF-beta/TAB1-mediated p38MAPK activation in human amnion epithelial cells †

doi: 10.1093/biolre/ioy135

Figure Lengend Snippet: Inhibition of ASK1 and MKK3/6 does not inhibit p38MAPK activation. (a) mRNA levels of ASK1 showed siRNA to ASK1 decreases its expression by 89% (P < .0001) compared to NT siRNA. A two-tailed t-test was used to test statistical significance. (b) AECs treated with CSE induced P-p38MAPK, but treatment with siRNA to ASK1 and CSE did not reduce p38MAPK phosphorylation. (c) mRNA levels of MKK3/6 showed siRNA to MKK3/6 decreases its expression by 90% (P < .0002) compared to NT siRNA. A two-tailed t-test was used to test statistical significance. (d) AECs treated with CSE induced P-p38mapk, as seen previously, but treatment with siRNA to MKK3/6 and CSE did not reduce P38MAPK phosphorylation. (e) Quantitative densitometry (b and d) of P-p38MAPK over total p38MAPK shows a significant increase of activated p38MAPK when treated with CSE (P < .003); however, its phosphorylation is not inhibited when treated with siRNA to ASK1 or MKK3/6 and CSE.

Article Snippet: The following antihuman antibodies were used for western blot: ASK1 (1:750, Cell Signaling, Danvers, MA), TRX (1:1000, Abcam, Cambridge, United Kingdom), TAB1 (1:1000, R&D, Minneapolis, MN), P-TAB1 (1:800, Milipore, Thr431, Burlington, MA), P-p38MAPK (1:300, Cell Signaling, T180/Y182), p38MAPK (1:1000, Cell Signaling) (Table ). table ft1 table-wrap mode="anchored" t5 Table 3. caption a7 Protein Name Catalog Number Company Dilution Method ASK1 D11C9 Cell Signaling 1:750 Western Blot TRX Ab16965 Abcam 1:1000 Western Blot TAB1 AF3578 R&D 1:1000 Western Blot P-TAB1(Thr431) 06–1334 Milipore 1:800 Western Blot P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Western Blot p38 MAPK 9212S Cell Signaling 1:1000 Western Blot Β-Actin A5441 Sigma-Aldrich 1:20,000 Western Blot Cytokeratin-18 Ab668 Abcam 1:300 Immunofluorescence ASK1 Ab45178 Abcam 1:250 Immunofluorescence TRX Ab16965 Abcam 1:500 Immunofluorescence P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Immunofluorescence TAK1 Sc-7967 Santa Cruz 1:300 Immunofluorescence TAB1 AF3578 R&D 1:300 Immunofluorescence ASK1 D11C9 Cell Signaling 1:100 Immunoprecipitation TRX Ab16965 Abcam 1:100 Immunoprecipitation GSK3B mAb#9832 Cell Signaling 1:100 Immunoprecipitation Open in a separate window Antibodies used for the western blot.

Techniques: Inhibition, Activation Assay, Expressing, Two Tailed Test

Journal: Biology of Reproduction

Article Title: Oxidative stress-induced TGF-beta/TAB1-mediated p38MAPK activation in human amnion epithelial cells †

doi: 10.1093/biolre/ioy135

Figure Lengend Snippet: ASK1-Signolsome in AECs. (a) Immunofluorescence images of Cytokeritin-18 (CK-18) positive (green) AECs. Scale bar is set to 30 μM. (b) Western blot analysis—ASK1 (dimer 150 kDa top band, monomer 75 kDa lower band) and Trx are present in AECs, regardless of treatment. (c) Crystal violet stain (purple) was not expelled from AECs after CSE 1:10 treatment for 1 h. This was confirmed by plate reader analysis showing no difference in crystal violet intensity. (d) Cigarette smoke extract (CSE) treatment induced P-p38MAPK (P < .05), while NAC (P < .05) prevented its activation. (e) Densitometry of P-p38MAPK (1C) over total p38MAPK in CSE and CSE + NAC exposed AECs. A one-way ANOVA with the Tukey Multiple Comparisons Test was used to test statistical significance. (f) ASK1 pulldowns in columns 1, 2, 5, 6, 7, and 8 all show ASK1 present, but not Trx, in both control and CSE bead samples, while the Trx Pulldown in columns 3 and 4 show Trx present, but not ASK1, in both control and CSE bead samples.

Article Snippet: The following antihuman antibodies were used for western blot: ASK1 (1:750, Cell Signaling, Danvers, MA), TRX (1:1000, Abcam, Cambridge, United Kingdom), TAB1 (1:1000, R&D, Minneapolis, MN), P-TAB1 (1:800, Milipore, Thr431, Burlington, MA), P-p38MAPK (1:300, Cell Signaling, T180/Y182), p38MAPK (1:1000, Cell Signaling) (Table ). table ft1 table-wrap mode="anchored" t5 Table 3. caption a7 Protein Name Catalog Number Company Dilution Method ASK1 D11C9 Cell Signaling 1:750 Western Blot TRX Ab16965 Abcam 1:1000 Western Blot TAB1 AF3578 R&D 1:1000 Western Blot P-TAB1(Thr431) 06–1334 Milipore 1:800 Western Blot P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Western Blot p38 MAPK 9212S Cell Signaling 1:1000 Western Blot Β-Actin A5441 Sigma-Aldrich 1:20,000 Western Blot Cytokeratin-18 Ab668 Abcam 1:300 Immunofluorescence ASK1 Ab45178 Abcam 1:250 Immunofluorescence TRX Ab16965 Abcam 1:500 Immunofluorescence P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Immunofluorescence TAK1 Sc-7967 Santa Cruz 1:300 Immunofluorescence TAB1 AF3578 R&D 1:300 Immunofluorescence ASK1 D11C9 Cell Signaling 1:100 Immunoprecipitation TRX Ab16965 Abcam 1:100 Immunoprecipitation GSK3B mAb#9832 Cell Signaling 1:100 Immunoprecipitation Open in a separate window Antibodies used for the western blot.

Techniques: Immunofluorescence, Western Blot, Staining, Activation Assay

Journal: Biology of Reproduction

Article Title: Oxidative stress-induced TGF-beta/TAB1-mediated p38MAPK activation in human amnion epithelial cells †

doi: 10.1093/biolre/ioy135

Figure Lengend Snippet: Immunofluorescence staining and Western blots of ASK1 and Trx in AECs. (a) Confocal microscopy was used to analyze the ratio of ASK1 and Trx in AECs. Trx (red) and ASK1 (green) staining can be seen in the cytoplasm and nuclei of control AECs. White lines represent regions of interest to look for ASK1-Trx colocalization. Scale bar is set to 10 μM. (b) Control and CSE graphs of ASK1-Trx showing no colocalization and ASK1 in the nucleus. (c) Western blot analysis of nuclear and cytoplasmic lysates confirms ASK1 (dimer 150 kDa top band, monomer 75 kDa lower band) in the nucleus and Trx in the cytoplasm.

Article Snippet: The following antihuman antibodies were used for western blot: ASK1 (1:750, Cell Signaling, Danvers, MA), TRX (1:1000, Abcam, Cambridge, United Kingdom), TAB1 (1:1000, R&D, Minneapolis, MN), P-TAB1 (1:800, Milipore, Thr431, Burlington, MA), P-p38MAPK (1:300, Cell Signaling, T180/Y182), p38MAPK (1:1000, Cell Signaling) (Table ). table ft1 table-wrap mode="anchored" t5 Table 3. caption a7 Protein Name Catalog Number Company Dilution Method ASK1 D11C9 Cell Signaling 1:750 Western Blot TRX Ab16965 Abcam 1:1000 Western Blot TAB1 AF3578 R&D 1:1000 Western Blot P-TAB1(Thr431) 06–1334 Milipore 1:800 Western Blot P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Western Blot p38 MAPK 9212S Cell Signaling 1:1000 Western Blot Β-Actin A5441 Sigma-Aldrich 1:20,000 Western Blot Cytokeratin-18 Ab668 Abcam 1:300 Immunofluorescence ASK1 Ab45178 Abcam 1:250 Immunofluorescence TRX Ab16965 Abcam 1:500 Immunofluorescence P-p38 MAPK (T180/y182) 9211L Cell Signaling 1:300 Immunofluorescence TAK1 Sc-7967 Santa Cruz 1:300 Immunofluorescence TAB1 AF3578 R&D 1:300 Immunofluorescence ASK1 D11C9 Cell Signaling 1:100 Immunoprecipitation TRX Ab16965 Abcam 1:100 Immunoprecipitation GSK3B mAb#9832 Cell Signaling 1:100 Immunoprecipitation Open in a separate window Antibodies used for the western blot.

Techniques: Immunofluorescence, Staining, Western Blot, Confocal Microscopy