Journal: Cell Communication and Signaling : CCS

Article Title: Bone morphogenetic protein receptor 2 signaling mediates mitochondrial Ca 2+ transport through its regulation of TAK1 splice variant

doi: 10.1186/s12964-025-02609-x

Figure Lengend Snippet: BMPR2 regulates TAK1-d splice variant to mediate mitochondrial Ca 2+ transport. A Western blot analysis of NSCLC and leukemia cell lines. B Western blot analysis of cells transfected with TAB1 cDNA or control vector after 24 h. C-D Western blot analysis of cells treated with JL189 for ( C ) 2 h or for ( D ) 4 h. E Western blot analysis of Calu-1 cells transfected with BMPR2 and control siRNA for 48 h. Graphs depict changes in the mean phosphorylation of TAK1 Thr184/Thr187 and TAK1 Ser412 expression of 3 independent biological replicates. F Western blot analysis of A549 WT and BMPR2 KO cells. G-H Western blot analysis of cells treated with BMPR1 receptor inhibitors. THP1 cells were treated for 4 h. Graphs represent normalized pTAK1 expression of (G) 3 independent biological replicates and ( H ) 4 independent biological replicates. I Western blot analysis demonstrating TAK1 splice variants using an antibody recognizing the N-terminal end of TAK1. The top is the immunoblot with a short film exposure and bottom immunoblot is with longer film exposure. J Western blot of THP1 cells treated with JL189 for 6 h. Graph represents normalized values of 2 independent biological replicates. K Western blot of cells transfected with TAB1 or control plasmid vectors for 24 h. Graph represents normalized values of 2 independent biological replicates. L-N Flow cytometry analysis of the fluorescence of Rhod-2AM of cells treated with TAK1 inhibitor 5Z-7-OXO 2 µM or 4 µM for 2 h. Graphs represent mean fluorescence of 3 independent biological replicates. O Western blot analysis of cells transfected with control or TAB1 siRNA for 48 h. Graph represents the mean percent change compared to control in normalized pTAK1 expression of 3 independent biological replicates. P-Q Flow cytometry analysis of the fluorescence of Rhod-2AM of cells transfected with TAB1 siRNA or ( M ) TAB1 cDNA. The graphs represent the mean Rhod-2AM fluorescence intensity of 3 (TAB1 siRNA) and 4 (TAB1 cDNA) independent biological replicates. Graphs show standard errors and p values represent the paired Student’s t test, assuming unequal variances

Article Snippet: BMPR2 (Cat. No. SR319490) and TAB1 (Cat. No. SR307092) siRNA were purchased from OriGene (Rockville, MD, USA).

Techniques: Variant Assay, Western Blot, Transfection, Control, Plasmid Preparation, Phospho-proteomics, Expressing, Flow Cytometry, Fluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Bone morphogenetic protein receptor 2 signaling mediates mitochondrial Ca 2+ transport through its regulation of TAK1 splice variant

doi: 10.1186/s12964-025-02609-x

Figure Lengend Snippet: BMPR2 regulation of mitochondrial Ca 2+ transport through the MCU effects mitochondrial bioenergetics and cell survival. A Western blot analysis of Jurkat and MDA-MB-231 WT and MCU KO cells. B Flow cytometry analysis of the fluorescence of Rhod-2AM of Jurkat WT and MCU KO cells treated with JL189 1.25 µM for 16 h. The graph represents the mean fluorescence of 4 independent biological replicates presented as percent change from its vehicle DMSO control. C Flow cytometry analysis of the fluorescence of Rhod-2AM of MDA-MB-231 WT and MCU KO cells treated with JL189 2.5 µM for 5 h. The graph represents the mean fluorescence of 2 independent biological replicates presented as percent change from its vehicle DMSO control. D-E Cell counts of ( D ) Jurkat and ( E ) MDA-231 WT and MCU KO cells treated with JL189 2.5 µM for 24 and/or 48 h. The graphs represent the mean percent number of dead cells of at least 4 independent biological replicates. F Flow cytometry analysis of the fluorescence of MitoTracker Green of MDA-MB-231 WT and MCU KO cells treated with JL189 2.5 µM for 16 h. The graph shows with mean fluorescence of 5 independent biological replicates presented as percent change from its vehicle DMSO control. G , I Western blot analysis of THP1 and H1299 transfected with TAB1 cDNA for 24 h done in duplicate. Graphs represent normalized TFAM1 and cytochrome b expression as the mean percent change from control of ( G ) 3 and ( I ) 2 independent biological replicates. H , J Western blot of THP1 and H1299 cells treated with 5Z-7-OXO for 2 h. Graphs represent normalized TFAM1 and cytochrome b expression as the mean percent change from control. H 3 independent biological replicates for TFAM1 and cytochrome b. J 3 independent biological replicates for TFAM1 and 2 independent biological replicates for cytochrome b. K Western blot of H1299 cells transfected with control or MCU siRNA for 48 h then treated with 5Z-7-OXO for 2 h. Graph represents the mean normalized TFAM1 expression of 3 independent biological replicates. L Western blot of H1299 tumor xenografts treated with vehicle or JL189 for 3 weeks. Graphs represent normalized values of 6 independent biological replicates. M Western blot analysis of immunoprecipitated (IP) TAB1-Flag or whole cell lysates of Calu-1 cells transfected with insertless cDNA or TAB1-Flag plasmids. Upper immunoblots show short film exposure and lower blot longer film exposure. N-O CellTiter-Glow assay was used to measure ATP levels. N Cells were treated with DMSO or JL189 2.5 µM for 4 h, or 1.5 µM oligomycin A for 50 min. O Cells were treated with DMSO or JL189 2.5 µM for 4 h. Graphs represent 3 independent biological replicates presented as the percent change from control. Graphs show standard errors and p values represent the paired Student’s t test, assuming unequal variances

Article Snippet: BMPR2 (Cat. No. SR319490) and TAB1 (Cat. No. SR307092) siRNA were purchased from OriGene (Rockville, MD, USA).

Techniques: Western Blot, Flow Cytometry, Fluorescence, Control, Transfection, Expressing, Immunoprecipitation

Journal: Cell reports

Article Title: A Tyrosine Switch on NEDD4-2 E3 Ligase Transmits GPCR Inflammatory Signaling

doi: 10.1016/j.celrep.2018.08.061

Figure Lengend Snippet:

Article Snippet: Rabbit anti-TAB1 , Cell signaling Technology , Cat# 3226S; RRID:AB_2140247.

Techniques: Recombinant, Sequencing, Negative Control, Software

Journal: Experimental and Therapeutic Medicine

Article Title: Combination of Sophora flavescens alkaloids and Panax quinquefolium saponins modulates different stages of experimental autoimmune myocarditis via the NF-κB and TGF-β1 pathways

doi: 10.3892/etm.2022.11507

Figure Lengend Snippet: Effects of KX on TAB1 and NF-κB signaling in the mouse myocardium. (A) Western blot analysis of TAB1, IKKα, IκB, p-IκB, NF-κB, p-NF-κB and TGF-β1 protein expression in EAM mice after 21 days. (B) Western blot quantification analysis (n=4 per group). (C) Immunohistochemistry analysis of NF-κB and TGF-β1 expression in EAM mice after 21 days. Scale bar, 100 µm. Arrows indicate positive expression of NF-κB or TGF-β1. (D) Immunohistochemical quantification of NF-κB and TGF-β1 expression in EAM mice after 21 days. n=10 per group. ** P<0.01 vs. Control. # P<0.05 and ## P<0.01 vs. EAM. TAB1, TGF-β activated kinase 1-binding protein 1; EAM, experimental autoimmune myocarditis; KX, combination of Sophora flavescens alkaloids (KuShen) and Panax quinquefolium saponins (XiYangShen).

Article Snippet: After blocking in 5% non-fat milk diluted in TBS-Tween 20 (0.1% Tween; CoWin Biosciences) in the dark at 4˚C overnight, the membranes were incubated with primary antibodies (Santa Cruz Biotechnology, Inc.) against activated kinase 1 (TAK1)-binding protein 1 (TAB1, cat. no. sc-166138; 1:100 dilution), NF-κB (cat. no. sc-8008; 1:200 dilution), phosphorylated (p)-NF-κB (cat. no. sc-136548; 1:200 dilution), IκB (cat. no. sc-74451; 1:100 dilution), p-IκB (cat. no. sc-8404; 1:200 dilution), IKKα (cat. no. sc-7606; 1:200 dilution), TGF-β1 (cat. no. sc-130348; 1:200 dilution), Smad2 (cat. no. sc-101153; 1:200 dilution), Smad4(cat. no. sc-7966; 1:200 dilution), Col Ⅰ (cat. no. sc-376350; 1:100 dilution) or GAPDH (cat. no. sc-365062; 1:100 dilution) for 2 h at 37˚C.

Techniques: Western Blot, Expressing, Immunohistochemistry, Immunohistochemical staining, Binding 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: 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: Production and function of TGF-beta in AECs. (a) ELISA for TGF-beta conducted on AEC supernatant that had been stimulated with CSE for 48 h. AECs showed a significantly increased production of TGF-beta when stimulated with CSE (P < .05) and NAC can inhibit this production (P < .05). A one-way ANOVA with the Tukey Multiple Comparisons Test was used to test statistical significance. (b) AECs treated with 2 ng/mL of TGF-beta for up to 1 h caused activation of p38MAPK (two-fold) compared to control treated AECs. (c) AECs treated with CSE for 1 h significantly induced p38MAPK activation (P < 0.0001) while TGF receptor antagonist prevented p38MAPK activation (P = 0.03). (d) ELISA for TGF-beta conducted on amniotic fluid of TNIL or TL deliveries showed significant expression of TGF-beta at term (P < .05). A one-tailed t-test was used to test statistical significance. (e) Immunofluorescence colocalized TAK (red) and TAB1 (green) inside control treated AECs. White lines represent regions of interest to look for TAK1-TAB1 colocalization. Both cells showed overlapping line graphs of TAK1-TAB1 documenting colocalization within the cytoplasm. Scale bar is set to 30 μM. (f) Western blot analysis confirmed that TAB1 can be significantly (P = .047) activated by CSE and antioxidant NAC can significantly prevent this activation (P < = .057). (g) Quantitative densitometry of P-TAB1 over actin for panel f showing statistical significance. A one-way ANOVA with the Tukey Multiple Comparisons Test was used to test statistical significance.

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: Enzyme-linked Immunosorbent Assay, Activation Assay, Expressing, One-tailed Test, Immunofluorescence, Western Blot

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: OS induces a TGF-beta–TAB1-dependent activation of p38MAPK. (a) mRNA levels of TAB1 showed siRNA to TAB1 decrease its expression by 91% (P < .0001) compared to NT siRNA. (b) AECs treated with CSE and siRNA to TAB1 significantly reduce p38MAPK phosphorylation (P = .0295). A two-tailed t-test was used to test statistical significance. (c) Quantitative densitometry (b) of P-p38MAPK over total p38MAPK shows a significant decrease of activated p38MAPK phosphorylation when treated with siRNA to TAB1 and CSE (P = .0295). A two-tailed t-test was used to test statistical significance.

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: Activation Assay, Expressing, Two Tailed Test