Journal: Aging (Albany NY)

Article Title: CUEDC1 inhibits epithelial-mesenchymal transition via the TβRI/Smad signaling pathway and suppresses tumor progression in non-small cell lung cancer

doi: 10.18632/aging.103329

Figure Lengend Snippet: CUEDC1 regulates TβRI/Smad signaling pathway. ( A , B ) Western blotting analysis of components of the TβR I/Smad signaling pathway and Snail in NSCLC cells. ( C ) The migration and invasion ability of shRNA/CUEDC1 or negative control in H1299 and A549 cells, with or without SB431542 treatment, was detected using transwell assays. P values were calculated using Student’s t -test. ( D ) H1299 and A549 cells stably transfected with CUEDC1-shRNA or empty vector were treated with 10 μM SB431542 for 24 h, and EMT markers was determined by immunoblotting. ( E ) CUEDC1-knockdown A549 cells or negative control A549 cells were treated with or without SB431542 for 24 h. Cells were immunostained with the indicated antibodies against EMT marker proteins. Experiments were performed at least three times. The data are expressed as the mean ± SEM; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: In this study, 10 μM SB431542 (HY-10431, MCE), 50 μg/ml CHX (HY-12320, MCE) and 10 μM MG132 (HY-13259, MCE) were used.

Techniques: Western Blot, Migration, shRNA, Negative Control, Stable Transfection, Transfection, Plasmid Preparation, Knockdown, Marker

Journal: The Journal of investigative dermatology

Article Title: Thrombospondin-1 Is a Major Activator of TGF-β Signaling in Recessive Dystrophic Epidermolysis Bullosa Fibroblasts.

doi: 10.1016/j.jid.2019.01.011

Figure Lengend Snippet: Figure 2. Phosphorylated smad3 levels are reduced after TSP1 siRNA knockdown or C7 overexpression in RDEB fibroblasts. (a) Transient siRNA knockdown of TSP1 (KD) down-regulates TGF-b signaling in RDEB fibroblasts compared with mock transfection (M) or scrambled siRNA control (C). The TGF-bR1 inhibitor SB431542 is used as a positive control (I). Graph shows mean ( standard deviation) quantitation of phosphorylated smad3 (PO4- SMAD3) levels relative to GAPDH for six independent experiments. Normal human fibroblasts are represented by Br43F, Br46F, and Br48F. RDEBSCC fibroblasts are represented by RDEB70F and RDEB71F, and RDEB fibroblasts are represented by RDEB75F, RDEB81F, and RDEB84F. (b) Overexpression of C7 significantly down-regulates TGF-b signaling in RDEB fibroblasts (RDEB70SCCF) but not in normal human fibroblasts Br43F. Graph shows mean ( standard deviation) quantitation of phosphorylated smad3 (PO4- SMAD3) levels relative to GAPDH for three independent experiments. *P < 0.05, **P < 0.005. C7, collagen VII; NHF, normal human fibroblast; RDEB, recessive dystrophic epidermolysis bullosa; siRNA, small interfering RNA; TGF, transforming growth factor.

Article Snippet: TGF-b and peptide treatment of fibroblast-derived ECM Matrices had the following treatments: control peptide SLLK, (AS-60875; AnaSpec, Fremont, CA) or LSKL thrombospondin 1 inhibitor (AS-60877, Anaspec) at 20 mmol/L; SB431542 TGF-beta inhibitor 1 mmol/L (S-1067; Selleckchem, Houston, TX); TGF-b ligand (240-B-002; R&D Systems, Minneapolis, MN) at 2 ng/ml.

Techniques: Knockdown, Over Expression, Transfection, Control, Positive Control, Standard Deviation, Quantitation Assay, Small Interfering RNA

Journal: Developmental biology

Article Title: An organizing role for the TGF-β signaling pathway in axes formation of the annelid Capitella teleta.

doi: 10.1016/j.ydbio.2018.01.004

Figure Lengend Snippet: Fig. 3. Early expression of components in the Activin/Nodal pathway (A) Schematic depicts components of the Activin/Nodal signaling pathway and highlights inhibition of the phosphorylation of receptor ALK4/5/7 by the chemical inhibitor SB431542. (B-D′′) 16 cell stage embryos. Images with the same letter correspond to images of a single embryo. (B, C, D) Merged confocal stack images labeled with anti-histone antibody to label nuclei. (B′, C′, D′) Merged confocal stack images showing expression of Ct-ALK4/5/7, Ct-Activin Receptor 2 (Ct-ACVR2), and Ct-SMAD2/3 by fluorescent in situ hybridization. (B′′, C′′, D′′) are merged confocal stack images of embryos labeled for both nuclei and gene expression.

Article Snippet: For inhibition of the Activin/Nodal signaling pathway, SB431542 (Santa Cruz; Cat No: SC-204265A), a chemical inhibitor that prevents the phosphorylation of ALK4, ALK5, ALK7 Activin/Nodal type 1 receptors, was used.

Techniques: Expressing, Inhibition, Phospho-proteomics, Labeling, In Situ Hybridization, Gene Expression

Journal: Developmental biology

Article Title: An organizing role for the TGF-β signaling pathway in axes formation of the annelid Capitella teleta.

doi: 10.1016/j.ydbio.2018.01.004

Figure Lengend Snippet: Fig. 4. Early exposure to SB431542 leads to abnormal larval morphology. Each row contains images of the same stage 6 larva raised in sea water or treated at a specified embryonic cell stage with either 0.4% DMSO as a control or 40 μM SB431542. Each column depicts either a DIC image (A-E) or merged confocal stack labeled for nuclei with Hoechst (A′- E′), cilia and neurons with anti-acetylated tubulin (A′′-E′′), or actin filaments with phalloidin (A′′′- E′′′). Images with the same letter correspond to images of a single animal. Figures A-A′′′ are images of a normal larva raised in sea water. Figures B-B′′′ are larvae treated with 0.4% DMSO during the 4–32 cell stage. Figures C-C′′′ are images of a larva treated with 40 μM SB431542 during the 4–32 cell stage. Figures D-D′′′ are images of control larvae treated with 0.4% DMSO during the 32–256 cell stage. Figures E-E′′′ are images of a larva treated with 40 μM SB431542 during the 32–256 cell stage. Open arrowhead, acetylated tubulin positive neurons; asterisk, position of the mouth; br, brain; cb, ciliary band; cm, circumferential muscle fibers; cn, clusters of nuclei; ec, ectodermal clearing; fg, foregut; lm, longitudinal muscle fibers; nt, neurotroch; pt, prototroch; tt, telotroch; vnc, ventral nerve cord.

Article Snippet: For inhibition of the Activin/Nodal signaling pathway, SB431542 (Santa Cruz; Cat No: SC-204265A), a chemical inhibitor that prevents the phosphorylation of ALK4, ALK5, ALK7 Activin/Nodal type 1 receptors, was used.

Techniques: Control, Labeling

Journal: Developmental biology

Article Title: An organizing role for the TGF-β signaling pathway in axes formation of the annelid Capitella teleta.

doi: 10.1016/j.ydbio.2018.01.004

Figure Lengend Snippet: Fig. 5. Analysis of anterior features following exposure to SB431542. Each row contains images of a stage 6 larva in anterior view. Each column contains either a merged confocal stack image of larvae labeled for nuclei and acetylated tubulin or a DIC image depicting the expression of the axial markers, CapI-elav1 and Ct-ADMP. Figures A-C are images of control larvae treated with 0.4% DMSO (a representative control phenotype). Figures D-F are images of larvae treated with 40 μM SB431542 during the 4–32 cell stage. Figures G-I are images of larvae treated with 40 μM SB431542 during the 32–256 cell stage. Arrowhead, acetylated tubulin positive sensory cells; black arrows, expression domains; white bracket, position of bilateral brain lobes.

Article Snippet: For inhibition of the Activin/Nodal signaling pathway, SB431542 (Santa Cruz; Cat No: SC-204265A), a chemical inhibitor that prevents the phosphorylation of ALK4, ALK5, ALK7 Activin/Nodal type 1 receptors, was used.

Techniques: Labeling, Expressing, Control

Journal: Developmental biology

Article Title: An organizing role for the TGF-β signaling pathway in axes formation of the annelid Capitella teleta.

doi: 10.1016/j.ydbio.2018.01.004

Figure Lengend Snippet: Fig. 6. Early exposure to SB431542 leads to abnormal expression of larval axial markers. Panels show expression of CapI-elav1, CapI-foxA, Ct-ADMP, and Ct-fox AB following in situ hybridization. A-D are larvae resulting from treatment with DMSO during the 4–32 cell stage (selected as a representative of control phenotype). E-H′ are larvae resulting from treatment with SB431542 during the 4–32 cell stage. I-L are larvae treated with SB431542 during the 32–256 cell stage. Specimens used to visualize Ct-fox AB expression were exposed to 50 μM SB431542 or 0.5% DMSO. All other specimens were exposed to 40 μM SB431542 or 0.4% DMSO. H & H′ are images of the same larva in different orientations (H, lateral view; H′ posterior view). All images are taken with DIC optics. Arrows indicate expression domains; asterisk indicates the position of the mouth; br, brain; dashed lines indicate position of ciliary band; fg, foregut; ve, ventral ectoderm; vnc, ventral nerve cord.

Article Snippet: For inhibition of the Activin/Nodal signaling pathway, SB431542 (Santa Cruz; Cat No: SC-204265A), a chemical inhibitor that prevents the phosphorylation of ALK4, ALK5, ALK7 Activin/Nodal type 1 receptors, was used.

Techniques: Expressing, In Situ Hybridization, Control

Journal: Developmental biology

Article Title: An organizing role for the TGF-β signaling pathway in axes formation of the annelid Capitella teleta.

doi: 10.1016/j.ydbio.2018.01.004

Figure Lengend Snippet: Fig. 7. Persistence of 2d lineage and its contribution to a reduced larval trunk Figures A-C depict control animals treated with 0.4% DMSO at the 4–32 cell stage. (A) Organizer cell 2d is the largest cell in the second quartet. (B) Daughter cells of 2d, 2d1 and 2d2. (C) Larva resulting from 2d microinjection with DiI showing trunk ectoderm and pygidium descendants (pink). Figures D-F depict animals treated with 40 μM SB431542 at the 4–32 cell stage. (D) In the presence of drug, cell 2d is born with its characteristic large size. (E) 2d daughter cells, 2d1 and 2d2 are born. (F) Larva resulting from 2d microinjection with DiI in drug treated animals. 2d generates a reduced trunk (pink). Larvae were imaged live in Figures C and F. Cb, ciliary band; cyan, nuclei; pt, prototroch; tt, telotroch; white, phalloidin labeling; white dotted line, ciliary band position.

Article Snippet: For inhibition of the Activin/Nodal signaling pathway, SB431542 (Santa Cruz; Cat No: SC-204265A), a chemical inhibitor that prevents the phosphorylation of ALK4, ALK5, ALK7 Activin/Nodal type 1 receptors, was used.

Techniques: Control, Microinjection, Labeling

Journal: Cellular Oncology

Article Title: Upregulation of limb-bud and heart (LBH) drives liver cancer progression by interacting with the oncoprotein Mortalin

doi: 10.1007/s13402-026-01228-z

Figure Lengend Snippet: LBH is transcriptionally upregulated by TGF-β in liver cancer cells. (A) Heatmap displaying the common 474 DEGs whose expressions were altered upon treatment with 100 pM of TGF-β1 for 4 h in three different HCC cell lines, i.e. MHCC97H, MHCC97L and HCCLM3. (B) Heatmap classification displaying some typical upregulated genes in response to TGF-β1 administration in HCC cell lines. (C-H) After being treated with 100 pM of TGF-β1 (C-E) or 10 µM of SB431542 (F-H) for the indicated periods, HCCLM3, HLE and SMMC-7721 cells were harvested for RNA purification and gene expression analyses through qPCR. (I-J) HCCLM3, HLE cells transfected with a control siRNA (NS) or those targeting Smad2/3 were treated with 100 pM of TGF-β1 for 6 h. Then the expression levels of Smad2, Smad3 and LBH were analyzed by qPCR. (K) Gene track view of Smad2/3, H3K4me3, and H3K27ac at the LBH promoter from the Cistrome Data database (top), and schematic diagram of ChIP primers designed within and near the LBH promoter region (bottom). HCCLM3 cells were treated with 2.5 ng/ml TGF-β for 24 h. ChIP assays were then performed to examine the binding of Smad2/3 to the LBH promoter region, and the enriched DNA fragments were quantitatively analyzed by qPCR. qPCR experiments described above were analyzed by one-way ANOVA followed by Tukey’s or Dunnett’s post hoc test. Statistical significance: * p < 0.05, ** p < 0.01 and *** p < 0.001; ns, no significance. Data were shown as mean ± SD

Article Snippet: Recombinant human TGF-β1 protein was obtained from R&D Systems Inc., and the small molecule inhibitor SB431542 was purchased from MedChemExpress (MCE, USA).

Techniques: Purification, Gene Expression, Transfection, Control, Expressing, Binding Assay