Journal: Cell Death Discovery

Article Title: LHPP suppresses colorectal cancer cell migration and invasion in vitro and in vivo by inhibiting Smad3 phosphorylation in the TGF-β pathway

doi: 10.1038/s41420-021-00657-z

Figure Lengend Snippet: a Hierarchical clustering of genes which were differentially expressed in Caco2 LHPP overexpression group and negative control group; The cut-off values were | Fold change | ≥1.2 and FDR < 0.05. b Volcano plot: red points mean genes with statistical difference at least 1.2-fold change and FDR < 0.05. c Ingenuity pathway analysis. The orange rectangle means activated cell pathway and the blue rectangle means inactivated cell pathway. Positive z score >2.0 was regarded as highly activated. Conversely, a negative z score < −2.0 was regarded as highly inhibited; TGF-β/Smad signaling pathway was inactivated after upregulating the expression of LHPP in the Ccaco2 cell line. Pivotal molecules of TGF-β/Smad signaling pathway, including Smad3, p-Smad3, Smad4, Smad7, Jun, and Fos, were evaluated in Sw480 ( d ) and Caco2 and HT-29 cell lines ( e ) using western blot assay. f – i Relative expression levels of these proteins were calculated using Image J and Graphpad Prism 6 software. j Cell nuclear proteins, such as p-Smad3, Smad4, Snail, Slug, and Twist1 were also examined using western blot, Histon-H3 was served as control. The quantification results were presented in k to n . * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Next, the TGF-β/Smad inhibitor-SIS3-(MCE, USA, HY-13013) was added to cancer cells.

Techniques: Over Expression, Negative Control, Expressing, Western Blot, Software, Control

Journal: Cell Death Discovery

Article Title: LHPP suppresses colorectal cancer cell migration and invasion in vitro and in vivo by inhibiting Smad3 phosphorylation in the TGF-β pathway

doi: 10.1038/s41420-021-00657-z

Figure Lengend Snippet: a , b Expression level of p-Smad3 was evaluated by using immunofluorescence assay after TGF-β1 stimulation for 24 h, cell nuclei were counterstained with DAPI and observed under a fluorescence microscope (scale bar: 25 μm). c , d In response to TGF-β1 stimulation, key molecules involved in the TGF-β/Smad classical signaling pathway were determined using western blot assay. e , f Relative expression levels of these proteins were calculated using Image J and Graphpad Prism 6 software. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Next, the TGF-β/Smad inhibitor-SIS3-(MCE, USA, HY-13013) was added to cancer cells.

Techniques: Expressing, Immunofluorescence, Fluorescence, Microscopy, Western Blot, Software

Journal: Cellular and molecular biology (Noisy-le-Grand, France)

Article Title: Erythropoietin relieves neuronal apoptosis in epilepsy rats via TGF-β/Smad signaling pathway.

doi: 10.14715/cmb/2023.69.10.35

Figure Lengend Snippet: Figure 2. Impacts of different treatments on TGF-β/Smad signaling in the of rats detected via WB. (A) The expression of TGF-β1 in PTZ + rHuEPO group was remarkably higher than that in other groups (P<0.01), while it was distinctly lower in PTZ + SB431542 group than that in other groups (P<0.01). (B) PTZ + rHuEPO group had an higher expression of p-smad3 than other groups (P<0.01), while PTZ + SB431542 group manifested a markedly lower p-smad3 expres sion than other groups (P<0.05). **P<0.01: PTZ + rHuEPO group vs. other groups, #P<0.05: PTZ + SB431542 group vs. other groups, ##P<0.01: PTZ + SB431542 group vs. other groups.

Article Snippet: Cellular and Molecular Biology E-ISSN : 1165-158X / P-ISSN : 0145-5680 www.cellmolbiol.org USA), pentylenetetrazol (PTZ) and dimethyl sulfoxide (DMSO) (Sigma, St. Louis, MO, USA), SB431542 (TGF-β/Smad inhibitor, Selleck, Houston, TX, USA), recombinant human EPO (rHuEPO), primary antibodies against TGF-β1, phosphorylated smad3 (p-smad3), tumor necrosis factor receptor 1 (TNFR1), caspase-3 and β-Actin and horseradish peroxidase (HRP)-labeled secondary antibodies (Abcam, Cambridge, MA, USA), and one-step terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) apoptosis assay kit (Beyotime Biotechnology, Shanghai, China).

Techniques: Expressing

Journal: Cellular and molecular biology (Noisy-le-Grand, France)

Article Title: Erythropoietin relieves neuronal apoptosis in epilepsy rats via TGF-β/Smad signaling pathway.

doi: 10.14715/cmb/2023.69.10.35

Figure Lengend Snippet: Figure 3. Impacts of different treatments on apoptotic proteins in the hippocampus of rats detected via WB. (A) The expression of apop totic protein TNFR1 was increased remarkably after the modeling using PTZ compared with that in control group (P<0.01), and it was down-regulated evidently after injection with rHuEPO following the modeling using PTZ (P<0.01). However, the down-regulatory effect of rHuEPO on TNFR1 expression could be repressed markedly by SB431542 (P<0.01). (B) In comparison with that in control group, the expression of apoptotic protein caspase-3 was enhanced notably after the modeling with PTZ (P<0.01), the injection with rHuEPO after the modeling using PTZ could prominently down-regulate the caspase-3 expression (P<0.01), and SB431542 could distinctly inhibit the down-regulatory effect of rHuEPO on caspase-3 expression (P<0.01). **P<0.01: PTZ + rHuEPO group vs. other groups, #P<0.05: PTZ + SB431542 group vs. other groups, ##P<0.01: PTZ + rHuEPO + SB431542 group vs. other groups.

Article Snippet: Cellular and Molecular Biology E-ISSN : 1165-158X / P-ISSN : 0145-5680 www.cellmolbiol.org USA), pentylenetetrazol (PTZ) and dimethyl sulfoxide (DMSO) (Sigma, St. Louis, MO, USA), SB431542 (TGF-β/Smad inhibitor, Selleck, Houston, TX, USA), recombinant human EPO (rHuEPO), primary antibodies against TGF-β1, phosphorylated smad3 (p-smad3), tumor necrosis factor receptor 1 (TNFR1), caspase-3 and β-Actin and horseradish peroxidase (HRP)-labeled secondary antibodies (Abcam, Cambridge, MA, USA), and one-step terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) apoptosis assay kit (Beyotime Biotechnology, Shanghai, China).

Techniques: Expressing, Control, Injection, Comparison

Journal: Cancer Management and Research

Article Title: Aberrant CEACAM19 expression is associated with metastatic phenotype in penile cancer

doi: 10.2147/CMAR.S192385

Figure Lengend Snippet: Notes: ( A ) The effect of SB431542 (SB) on p-Smad2/3 levels in Penl1 cells. Penl1 cells were treated with 10 µM SB for 24 hours. β-Actin served as a loading control. ( B ) The effect of SB431542 on wound healing of Penl1 cells. Three different areas in each assay were chosen to measure the distances of migrating cells to the origin of the wound. * P <0.05, Control vs SB. ( C ) The effect of SB431542 on cell invasion of Penl1 cells. Transwell invasion assay was conducted to evaluate cell migration and invasion capability. * P <0.05, Control vs SB. ( D ) Inhibition on TGF-β/Smad2/3 activity by SB431542 reduced MMP2/9 secretion in Penl1 cells. All experiments were performed three times, and data are presented as mean ± SD. values. * P <0.05, Control vs SB.

Article Snippet: As shown in , p-Smad2/3 level was potently reduced by TGF-β/Smad2/3 pathway inhibitor SB431542 (Cell Signaling).

Techniques: Control, Transwell Invasion Assay, Migration, Inhibition, Activity Assay