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PeproTech human tgfβ3
Human Tgfβ3, supplied by PeproTech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 1 article reviews

human tgfβ3 - by Bioz Stars, 2026-04

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TGFβ3 potentiates palbociclib anti-tumor effect in vivo. a mRNA expression levels of TGFB1, TGFB2 and TGFB3 in SUM159PT following TGFB3 -specific overexpression using CRISPR activation (CRISPR/dCas9 SAM) ( n = 3). Data are represented as mean ± standard deviation (SD). Significance was calculated using two-sided, unpaired t-test, p -value * < 0.05. b Mice from control (lentiSAMv2) or TGFB3 -overexpressing (TGFB3g2 SAM) groups ( n = 13) were each randomized into treatment groups (vehicle, n = 6; palbociclib, n = 7). I.p. injections of the vehicle treatment or a low dose of palbociclib (10 mg/kg) were administered until study endpoint. Data are represented as mean ± SD. c Reduction in tumor growth presented for each group treated with palbociclib, lentiSAMv2 or TGFB3g2 SAM, as compared to the same groups treated with the vehicle. Data are represented as mean, at each timepoint. d left Tumor volumes of individual mice in each group at study endpoint. right Tumor weights of individual mice in each group at study endpoint. Midlines at median. Significance was calculated using ordinary, one-way ANOVA with Tukey’s multiple comparisons test, p -value * < 0.05, ** < 0.01, *** < 0.001. e Average mRNA expression levels of TGFB3 in tumors derived from the vehicle-treated control mice ( n = 6) and the TGFB3 -overexpressing mice ( n = 6). Data are represented as mean ± SD. Significance was calculated using two-sided, unpaired t-test, p -value * < 0.05, ** < 0.01, *** < 0.001. f Protein levels of TGFB3 (60 kDa) in tumors derived from the vehicle-treated control mice ( n = 6) and the TGFB3 -overexpressing mice ( n = 6). g Spontaneous metastasis to the lungs was assessed. Lung nodules were counted and compared in lungs derived from the vehicle-treated control mice ( n = 7) and the TGFB3 -overexpressing mice ( n = 6). Data represent metastatic nodule count per pair of lungs per mouse. Midlines at median. Significance was calculated using nonparametric Mann–Whitney U-test, p -value * < 0.05, ** < 0.01, *** < 0.001. h The effect of TGFB3 CRISPR-mediated knockout on lung colonization was assessed. Data represent metastatic nodule count per pair of lungs per mouse. Midlines at median. i Schematic representation of the use of recTGFβ3 in combination with palbociclib. MDA-MB-231 TNBC cells were transplanted into the mammary fat pads of NSG mice. Tumors were palpable before mice were randomized into treatment groups: vehicle, n = 9; recTGFβ3, n = 8; palbociclib, n = 8, combo (recTGFβ3 + palbociclib), n = 9. j Average tumor volume was measured over time. Data are represented as mean ± SD. k Tumor volumes of individual mice in each group at study endpoint. Midlines at median. Significance was calculated using ordinary, one-way ANOVA with Tukey’s multiple comparisons test, p -value * < 0.05. l Quantification of Ki67-positive cells stained by immunohistochemistry in tumor tissues from all four groups. Data are represented as mean ± SD ( n = 3–4). Significance was calculated using two-sided, unpaired t-test, p -value * < 0.05. m Representative images of Ki67 staining in two tumors per group

Journal: Molecular Cancer

Article Title: Genome-wide in vivo CRISPR screen identifies TGFβ3 as actionable biomarker of palbociclib resistance in triple negative breast cancer

doi: 10.1186/s12943-024-02029-4

Figure Lengend Snippet: TGFβ3 potentiates palbociclib anti-tumor effect in vivo. a mRNA expression levels of TGFB1, TGFB2 and TGFB3 in SUM159PT following TGFB3 -specific overexpression using CRISPR activation (CRISPR/dCas9 SAM) ( n = 3). Data are represented as mean ± standard deviation (SD). Significance was calculated using two-sided, unpaired t-test, p -value * < 0.05. b Mice from control (lentiSAMv2) or TGFB3 -overexpressing (TGFB3g2 SAM) groups ( n = 13) were each randomized into treatment groups (vehicle, n = 6; palbociclib, n = 7). I.p. injections of the vehicle treatment or a low dose of palbociclib (10 mg/kg) were administered until study endpoint. Data are represented as mean ± SD. c Reduction in tumor growth presented for each group treated with palbociclib, lentiSAMv2 or TGFB3g2 SAM, as compared to the same groups treated with the vehicle. Data are represented as mean, at each timepoint. d left Tumor volumes of individual mice in each group at study endpoint. right Tumor weights of individual mice in each group at study endpoint. Midlines at median. Significance was calculated using ordinary, one-way ANOVA with Tukey’s multiple comparisons test, p -value * < 0.05, ** < 0.01, *** < 0.001. e Average mRNA expression levels of TGFB3 in tumors derived from the vehicle-treated control mice ( n = 6) and the TGFB3 -overexpressing mice ( n = 6). Data are represented as mean ± SD. Significance was calculated using two-sided, unpaired t-test, p -value * < 0.05, ** < 0.01, *** < 0.001. f Protein levels of TGFB3 (60 kDa) in tumors derived from the vehicle-treated control mice ( n = 6) and the TGFB3 -overexpressing mice ( n = 6). g Spontaneous metastasis to the lungs was assessed. Lung nodules were counted and compared in lungs derived from the vehicle-treated control mice ( n = 7) and the TGFB3 -overexpressing mice ( n = 6). Data represent metastatic nodule count per pair of lungs per mouse. Midlines at median. Significance was calculated using nonparametric Mann–Whitney U-test, p -value * < 0.05, ** < 0.01, *** < 0.001. h The effect of TGFB3 CRISPR-mediated knockout on lung colonization was assessed. Data represent metastatic nodule count per pair of lungs per mouse. Midlines at median. i Schematic representation of the use of recTGFβ3 in combination with palbociclib. MDA-MB-231 TNBC cells were transplanted into the mammary fat pads of NSG mice. Tumors were palpable before mice were randomized into treatment groups: vehicle, n = 9; recTGFβ3, n = 8; palbociclib, n = 8, combo (recTGFβ3 + palbociclib), n = 9. j Average tumor volume was measured over time. Data are represented as mean ± SD. k Tumor volumes of individual mice in each group at study endpoint. Midlines at median. Significance was calculated using ordinary, one-way ANOVA with Tukey’s multiple comparisons test, p -value * < 0.05. l Quantification of Ki67-positive cells stained by immunohistochemistry in tumor tissues from all four groups. Data are represented as mean ± SD ( n = 3–4). Significance was calculated using two-sided, unpaired t-test, p -value * < 0.05. m Representative images of Ki67 staining in two tumors per group

Article Snippet: Recombinant human TGFβ3 ligand (PeproTech, Inc, cat. no. 100-36E) was dissolved in 10 mM citric acid buffer with 0.1% BSA.

Techniques: In Vivo, Expressing, Over Expression, CRISPR, Activation Assay, Standard Deviation, Control, Derivative Assay, MANN-WHITNEY, Knock-Out, Staining, Immunohistochemistry

Combination of recombinant TGFβ3 and palbociclib synergistically inhibits TNBC cell proliferation in vitro. a Synergy between palbociclib and recTGFβ3 dose combinations was calculated based on four reference models (Bliss, HSA, Loewe, ZIP) using SynergyFinder in four TNBC cell lines (MDA-MB-231, SUM159PT, SUM229PE, 159-R). Synergy maps highlight areas of synergistic (red) or antagonistic (green) interactions between given concentrations of either agent. Grey boxes indicate the area of maximum synergy observed. Mean of a minimum of three independent replicate experiments for each cell line ( n ≥ 3). b ‘Overall synergy scores’ and ‘Most synergistic area scores’ presented for each drug matrix shown in a . Data are represented as score ± 95% confidence interval. c Dot plots show overall synergy scores (black) or most synergistic area scores (pink) for each cell line, with each dot representing the score obtained using the indicated reference model. Midlines represent median scores. Outer vertical lines correspond to minimum and maximum scores obtained. A zero ‘0’ score indicates no interaction between the two agents

Journal: Molecular Cancer

Article Title: Genome-wide in vivo CRISPR screen identifies TGFβ3 as actionable biomarker of palbociclib resistance in triple negative breast cancer

doi: 10.1186/s12943-024-02029-4

Figure Lengend Snippet: Combination of recombinant TGFβ3 and palbociclib synergistically inhibits TNBC cell proliferation in vitro. a Synergy between palbociclib and recTGFβ3 dose combinations was calculated based on four reference models (Bliss, HSA, Loewe, ZIP) using SynergyFinder in four TNBC cell lines (MDA-MB-231, SUM159PT, SUM229PE, 159-R). Synergy maps highlight areas of synergistic (red) or antagonistic (green) interactions between given concentrations of either agent. Grey boxes indicate the area of maximum synergy observed. Mean of a minimum of three independent replicate experiments for each cell line ( n ≥ 3). b ‘Overall synergy scores’ and ‘Most synergistic area scores’ presented for each drug matrix shown in a . Data are represented as score ± 95% confidence interval. c Dot plots show overall synergy scores (black) or most synergistic area scores (pink) for each cell line, with each dot representing the score obtained using the indicated reference model. Midlines represent median scores. Outer vertical lines correspond to minimum and maximum scores obtained. A zero ‘0’ score indicates no interaction between the two agents

Article Snippet: Recombinant human TGFβ3 ligand (PeproTech, Inc, cat. no. 100-36E) was dissolved in 10 mM citric acid buffer with 0.1% BSA.

Techniques: Recombinant, In Vitro

TGFβ3 synergizes with palbociclib in a p21-dependent way . a SUM159PT cells were treated with palbociclib (100 nM) for 2 h, 8 h, 16 h and 24 h and protein lysates were assessed for known CDK4/6i resistance markers (CDK4, cyclin D1, cyclin E1, Rb, phospho-Rb (S780)) by immunoblotting. Relative fold changes in protein levels, compared to untreated cells at each timepoint, were calculated ( n = 3). Data are represented as mean ± SD. Significance was calculated using two-sided, unpaired t-test, p -value * < 0.05, ** < 0.01, *** < 0.001. b SUM159PT (159) and 159-R cells were assessed for known CDK4/6i resistance markers, as well as p21, by immunoblotting. c top SUM159PT and 159-R cells were treated with recTGFβ3 (100 pM) for 24 h and resulting changes in known CDK4/6i resistance markers and p21 were measured by immunoblotting. bottom MDA-MB-231 (231) and SUM229PE (229) cells were treated with recTGFβ3 (200 pM) for 24-48 h and resulting changes in p21 were measured by immunoblotting. d SUM159PT cells were transduced with plasmids encoding control (scramble, scr), Smad2-specific, or Smad3-specific short hairpin RNAs (shRNA). Protein levels of p21 and total Smad2/3 were measured by immunoblotting. e SUM159PT cells were transduced with plasmids encoding control (scr) and p21-specific shRNA. Protein levels of p21 were measured by immunoblotting. f SUM159PT scr shRNA-infected or p21 shRNA-infected cells were treated with varying combinations of palbociclib and recTGFβ3 concentrations. Synergy between dose combinations was calculated using SynergyFinder. upper Synergy maps highlight areas of synergistic (red) or antagonistic (green) interactions between given concentrations of either agent. Grey boxes indicate the area of maximum synergy observed between given recTGFβ3 and palbociclib dose combinations. lower ‘Overall synergy scores’ and ‘Most synergistic area scores’ presented for each drug matrix shown above. Data are represented as score ± 95% confidence interval ( n = 3). Percentage variation in synergy score (score obtained in p21 shRNA cells/score obtained in scr shRNA cells) is also shown (red)

Journal: Molecular Cancer

Article Title: Genome-wide in vivo CRISPR screen identifies TGFβ3 as actionable biomarker of palbociclib resistance in triple negative breast cancer

doi: 10.1186/s12943-024-02029-4

Figure Lengend Snippet: TGFβ3 synergizes with palbociclib in a p21-dependent way . a SUM159PT cells were treated with palbociclib (100 nM) for 2 h, 8 h, 16 h and 24 h and protein lysates were assessed for known CDK4/6i resistance markers (CDK4, cyclin D1, cyclin E1, Rb, phospho-Rb (S780)) by immunoblotting. Relative fold changes in protein levels, compared to untreated cells at each timepoint, were calculated ( n = 3). Data are represented as mean ± SD. Significance was calculated using two-sided, unpaired t-test, p -value * < 0.05, ** < 0.01, *** < 0.001. b SUM159PT (159) and 159-R cells were assessed for known CDK4/6i resistance markers, as well as p21, by immunoblotting. c top SUM159PT and 159-R cells were treated with recTGFβ3 (100 pM) for 24 h and resulting changes in known CDK4/6i resistance markers and p21 were measured by immunoblotting. bottom MDA-MB-231 (231) and SUM229PE (229) cells were treated with recTGFβ3 (200 pM) for 24-48 h and resulting changes in p21 were measured by immunoblotting. d SUM159PT cells were transduced with plasmids encoding control (scramble, scr), Smad2-specific, or Smad3-specific short hairpin RNAs (shRNA). Protein levels of p21 and total Smad2/3 were measured by immunoblotting. e SUM159PT cells were transduced with plasmids encoding control (scr) and p21-specific shRNA. Protein levels of p21 were measured by immunoblotting. f SUM159PT scr shRNA-infected or p21 shRNA-infected cells were treated with varying combinations of palbociclib and recTGFβ3 concentrations. Synergy between dose combinations was calculated using SynergyFinder. upper Synergy maps highlight areas of synergistic (red) or antagonistic (green) interactions between given concentrations of either agent. Grey boxes indicate the area of maximum synergy observed between given recTGFβ3 and palbociclib dose combinations. lower ‘Overall synergy scores’ and ‘Most synergistic area scores’ presented for each drug matrix shown above. Data are represented as score ± 95% confidence interval ( n = 3). Percentage variation in synergy score (score obtained in p21 shRNA cells/score obtained in scr shRNA cells) is also shown (red)

Article Snippet: Recombinant human TGFβ3 ligand (PeproTech, Inc, cat. no. 100-36E) was dissolved in 10 mM citric acid buffer with 0.1% BSA.

Techniques: Western Blot, Transduction, Control, shRNA, Infection

Schematic diagram depicting TGFβ3-palbociclib synergy. a In the basal context, cells maintain a balance between active (green) and p21-bound, inactive (red) CDK/cyclin complexes. In the presence of palbociclib (orange capsule), CDK4/6 kinase activity is inactivated, and p21 (pink box) bound to CDK4 is released and preferentially displaced to CDK2. This inactivates CDK2/cyclin E complexes and leads to overall cell cycle arrest. b upper When cells undergo prolonged exposure to palbociclib, key cell cycle regulators (CDK4, cyclins D and E) are upregulated, while p21 expression is strongly inhibited. Some CDK/cyclin complexes are inactivated (red), but the overall imbalance in active CDK4/cyclinD1 and CDK2/cyclinE1 complexes (green) leads to decreased responsiveness of cells to palbociclib, acquired resistance to the drug, and continued cell cycling. lower When TGFβ3 is added in the presence of palbociclib, p21 expression levels are restored through TGFβ3 signalling. The increase in p21 by TGFβ3 synergizes with palbociclib’s mechanism of action, allowing for the inactivation of all remaining active CDK/cyclin complexes (red), and ultimately leading to cell cycle arrest

Journal: Molecular Cancer

Article Title: Genome-wide in vivo CRISPR screen identifies TGFβ3 as actionable biomarker of palbociclib resistance in triple negative breast cancer

doi: 10.1186/s12943-024-02029-4

Figure Lengend Snippet: Schematic diagram depicting TGFβ3-palbociclib synergy. a In the basal context, cells maintain a balance between active (green) and p21-bound, inactive (red) CDK/cyclin complexes. In the presence of palbociclib (orange capsule), CDK4/6 kinase activity is inactivated, and p21 (pink box) bound to CDK4 is released and preferentially displaced to CDK2. This inactivates CDK2/cyclin E complexes and leads to overall cell cycle arrest. b upper When cells undergo prolonged exposure to palbociclib, key cell cycle regulators (CDK4, cyclins D and E) are upregulated, while p21 expression is strongly inhibited. Some CDK/cyclin complexes are inactivated (red), but the overall imbalance in active CDK4/cyclinD1 and CDK2/cyclinE1 complexes (green) leads to decreased responsiveness of cells to palbociclib, acquired resistance to the drug, and continued cell cycling. lower When TGFβ3 is added in the presence of palbociclib, p21 expression levels are restored through TGFβ3 signalling. The increase in p21 by TGFβ3 synergizes with palbociclib’s mechanism of action, allowing for the inactivation of all remaining active CDK/cyclin complexes (red), and ultimately leading to cell cycle arrest

Article Snippet: Recombinant human TGFβ3 ligand (PeproTech, Inc, cat. no. 100-36E) was dissolved in 10 mM citric acid buffer with 0.1% BSA.

Techniques: Activity Assay, Expressing

Ovarian FF TGFβ3 levels are positively correlated with oocyte maturity. ( A ) The scatter plot and paired Student’s t -test of FF TGFβ3 levels between a large (preovulatory) leading follicle (diameter > 18 mm) and small (mid-antral) follicles. ( B ) The correlation of FF TGFβ3 levels and oocyte maturity was performed by receiver operating characteristic (ROC) curve analysis (* p < 0.05).

Journal: International Journal of Molecular Sciences

Article Title: The Presence of TGFβ3 in Human Ovarian Intrafollicular Fluid and Its Involvement in Thromboxane Generation in Follicular Granulosa Cells through a Canonical TGFβRI, Smad2/3 Signaling Pathway and COX-2 Induction

doi: 10.3390/ijms25105558

Figure Lengend Snippet: Ovarian FF TGFβ3 levels are positively correlated with oocyte maturity. ( A ) The scatter plot and paired Student’s t -test of FF TGFβ3 levels between a large (preovulatory) leading follicle (diameter > 18 mm) and small (mid-antral) follicles. ( B ) The correlation of FF TGFβ3 levels and oocyte maturity was performed by receiver operating characteristic (ROC) curve analysis (* p < 0.05).

Article Snippet: Human TGFβ3 was obtained from ProSpec-Tany TechnoGene Ltd. (East Brunswick, NJ, USA).

Techniques:

Logistic regression analysis of TGFβ3 levels and oocyte maturity in mid-antral and preovulatory follicle groups.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25105558

Figure Lengend Snippet: Logistic regression analysis of TGFβ3 levels and oocyte maturity in mid-antral and preovulatory follicle groups.

Article Snippet: Human TGFβ3 was obtained from ProSpec-Tany TechnoGene Ltd. (East Brunswick, NJ, USA).

Techniques:

TGFβ3 increases TXB 2 production in ( A ) human HO23 GCs and ( B ) primary cultured ovarian follicular GCs. GCs were treated with the indicated concentrations of TGFβ3 for 24 h. The culture media were collected and immediately analyzed for TXB 2 production by ELISA ( n = 3). * p < 0.05, ** p < 0.01, and **** p < 0.0001 versus vehicle only (control; TGFβ3: 0 ng/mL) and the exact p -values for all significant differences are shown in each bar.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25105558

Figure Lengend Snippet: TGFβ3 increases TXB 2 production in ( A ) human HO23 GCs and ( B ) primary cultured ovarian follicular GCs. GCs were treated with the indicated concentrations of TGFβ3 for 24 h. The culture media were collected and immediately analyzed for TXB 2 production by ELISA ( n = 3). * p < 0.05, ** p < 0.01, and **** p < 0.0001 versus vehicle only (control; TGFβ3: 0 ng/mL) and the exact p -values for all significant differences are shown in each bar.

Article Snippet: Human TGFβ3 was obtained from ProSpec-Tany TechnoGene Ltd. (East Brunswick, NJ, USA).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

TGFβ3 enhances COX-2 protein and mRNA expression in human HO23 GCs and primary cultured ovarian follicular GCs. ( A ) Human HO23 GCs or ( B ) human primary cultured ovarian follicular GCs were treated with the indicated concentrations of TGFβ3 for 24 h, and COX-1 and COX-2 protein expression were analyzed by Western blotting. ( C ) Human HO23 GCs were treated with the indicated concentrations of TGFβ3 for 4 h. At the end of incubation, cells were collected, total RNA was extracted, and the COXs and β-actin mRNA were analyzed by RT-PCR. Data are expressed as mean ± SEM ( n = 3). * p < 0.05 and ** p < 0.01 versus vehicle only (control; TGFβ3: 0 ng/mL) and the exact p -values for all significant differences are shown.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25105558

Figure Lengend Snippet: TGFβ3 enhances COX-2 protein and mRNA expression in human HO23 GCs and primary cultured ovarian follicular GCs. ( A ) Human HO23 GCs or ( B ) human primary cultured ovarian follicular GCs were treated with the indicated concentrations of TGFβ3 for 24 h, and COX-1 and COX-2 protein expression were analyzed by Western blotting. ( C ) Human HO23 GCs were treated with the indicated concentrations of TGFβ3 for 4 h. At the end of incubation, cells were collected, total RNA was extracted, and the COXs and β-actin mRNA were analyzed by RT-PCR. Data are expressed as mean ± SEM ( n = 3). * p < 0.05 and ** p < 0.01 versus vehicle only (control; TGFβ3: 0 ng/mL) and the exact p -values for all significant differences are shown.

Article Snippet: Human TGFβ3 was obtained from ProSpec-Tany TechnoGene Ltd. (East Brunswick, NJ, USA).

Techniques: Expressing, Cell Culture, Western Blot, Incubation, Reverse Transcription Polymerase Chain Reaction

Effect of pharmacological interventions on TGFβ3-induced TXB 2 generation and COX-2 expression. Human HO23 GCs were treated with vehicle or TGFβ3 (10 ng/mL) with or without the indicated inhibitor for 24 h. ( A ) The culture media were collected and immediately analyzed for TXB 2 production by ELISA. ( B ) The COX-1 and -2 expressions were analyzed by Western blotting and quantitated by densitometry ( n = 3–6). I: inhibitor. ## p < 0.01 versus basal level, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus TGFβ3 control (—, +vehicle) and the exact p -values for all significant differences are shown.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25105558

Figure Lengend Snippet: Effect of pharmacological interventions on TGFβ3-induced TXB 2 generation and COX-2 expression. Human HO23 GCs were treated with vehicle or TGFβ3 (10 ng/mL) with or without the indicated inhibitor for 24 h. ( A ) The culture media were collected and immediately analyzed for TXB 2 production by ELISA. ( B ) The COX-1 and -2 expressions were analyzed by Western blotting and quantitated by densitometry ( n = 3–6). I: inhibitor. ## p < 0.01 versus basal level, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus TGFβ3 control (—, +vehicle) and the exact p -values for all significant differences are shown.

Article Snippet: Human TGFβ3 was obtained from ProSpec-Tany TechnoGene Ltd. (East Brunswick, NJ, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

TGFβ3 induces Smad2/3 phosphorylation in GCs. Human HO23 GCs were treated with vehicle or TGFβ3 (10 ng/mL) for the indicated time intervals. The Smad2/3 phosphorylation and its total protein expression were analyzed by Western blotting and densitometry ( n = 4). * p < 0.05 compared to vehicle treatment at the corresponding time point (15 and 30 min) and the exact p -values for the significant differences are shown.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25105558

Figure Lengend Snippet: TGFβ3 induces Smad2/3 phosphorylation in GCs. Human HO23 GCs were treated with vehicle or TGFβ3 (10 ng/mL) for the indicated time intervals. The Smad2/3 phosphorylation and its total protein expression were analyzed by Western blotting and densitometry ( n = 4). * p < 0.05 compared to vehicle treatment at the corresponding time point (15 and 30 min) and the exact p -values for the significant differences are shown.

Article Snippet: Human TGFβ3 was obtained from ProSpec-Tany TechnoGene Ltd. (East Brunswick, NJ, USA).

Techniques: Expressing, Western Blot

$TGFβ3 induces Smad2/3 translocation from cytosol to the nucleus. Human HO23 GCs were treated with vehicle or TGFβ3 (10 ng/mL) for the indicated time intervals. ( A ) The cell lysates were fractionated as a cytosolic and nuclear fraction, and p-Smad2/3 and total Smad2/3 expressions in cytosolic and nuclear compartments were determined by Western blotting and densitometry ( n = 5). ** p < 0.01 and *** p < 0.001 versus its corresponding controls (vehicle treatment at 10, 30, and 60 min) and the exact p -values for the significant differences are shown. ( B ) The Smad2/3 translocation was assayed by immunofluorescence microscopy. Merge: an overlay of images of Smad2/3 staining with DAPI staining. Scale bar = 50 μm.$

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25105558

Figure Lengend Snippet: TGFβ3 induces Smad2/3 translocation from cytosol to the nucleus. Human HO23 GCs were treated with vehicle or TGFβ3 (10 ng/mL) for the indicated time intervals. ( A ) The cell lysates were fractionated as a cytosolic and nuclear fraction, and p-Smad2/3 and total Smad2/3 expressions in cytosolic and nuclear compartments were determined by Western blotting and densitometry ( n = 5). ** p < 0.01 and *** p < 0.001 versus its corresponding controls (vehicle treatment at 10, 30, and 60 min) and the exact p -values for the significant differences are shown. ( B ) The Smad2/3 translocation was assayed by immunofluorescence microscopy. Merge: an overlay of images of Smad2/3 staining with DAPI staining. Scale bar = 50 μm.

Article Snippet: Human TGFβ3 was obtained from ProSpec-Tany TechnoGene Ltd. (East Brunswick, NJ, USA).

Techniques: Translocation Assay, Western Blot, Immunofluorescence, Microscopy, Staining

SiRNA knockdown of COX-2 expression compromises TGFβ3-induced TXB 2 production. The HO23 GCs were transfected with control or COX-2 siRNA (150 or 300 nM) for 24 h and followed by the addition of vehicle or TGFβ3 (10 ng/mL) for 4 h. The cells were collected, ( A ) total RNA was extracted, and COX-2 mRNA expression was determined by RT-PCR, whereas ( B ) the culture media were collected and TGFβ3-induced TXB 2 production was measured by ELISA. * p < 0.05 and ** p < 0.01 versus basal (control or COX-2 siRNA transfection with vehicle treatment only) and the exact p -values for the significant differences are shown. NS: not significant.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25105558

Figure Lengend Snippet: SiRNA knockdown of COX-2 expression compromises TGFβ3-induced TXB 2 production. The HO23 GCs were transfected with control or COX-2 siRNA (150 or 300 nM) for 24 h and followed by the addition of vehicle or TGFβ3 (10 ng/mL) for 4 h. The cells were collected, ( A ) total RNA was extracted, and COX-2 mRNA expression was determined by RT-PCR, whereas ( B ) the culture media were collected and TGFβ3-induced TXB 2 production was measured by ELISA. * p < 0.05 and ** p < 0.01 versus basal (control or COX-2 siRNA transfection with vehicle treatment only) and the exact p -values for the significant differences are shown. NS: not significant.

Article Snippet: Human TGFβ3 was obtained from ProSpec-Tany TechnoGene Ltd. (East Brunswick, NJ, USA).

Techniques: Expressing, Transfection, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Proposed illustrated scheme for TGFβ3′s effects on follicular GCs. TGFβ3 binds TGFβ receptor II and I (RII and RI), leading to an activation of the canonical signaling pathway, including Smad2/3 phosphorylation and translocation to the cell nucleus. The activated Smad2/3 acts as a transcription factor to drive COX-2 mRNA and protein induction, which then triggers TXA 2 generation and secretion into extracellular space. The indicates a pharmacological and/or siRNA intervention used in this study.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25105558

Figure Lengend Snippet: Proposed illustrated scheme for TGFβ3′s effects on follicular GCs. TGFβ3 binds TGFβ receptor II and I (RII and RI), leading to an activation of the canonical signaling pathway, including Smad2/3 phosphorylation and translocation to the cell nucleus. The activated Smad2/3 acts as a transcription factor to drive COX-2 mRNA and protein induction, which then triggers TXA 2 generation and secretion into extracellular space. The indicates a pharmacological and/or siRNA intervention used in this study.

Article Snippet: Human TGFβ3 was obtained from ProSpec-Tany TechnoGene Ltd. (East Brunswick, NJ, USA).

Techniques: Activation Assay, Translocation Assay

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