primary antibodies against gata6  (R&D Systems)

Journal: Frontiers in Physiology

Article Title: Transcription factor GATA6 promotes migration of human coronary artery smooth muscle cells in vitro

doi: 10.3389/fphys.2022.1054819

Figure Lengend Snippet: GATA6 induces vascular smooth muscle cell differentiation. Human coronary artery smooth muscle cells were transduced with Ad-CMV-GATA6/Ad-CMV-null (A,B) . Expression of GATA6 and some smooth muscle markers are presented in (A) at mRNA level and (B) at protein level. (C) Relative mRNA expression of targets after transfection with GapmeRs against GATA6. Data are presented as mean ± SEM ( n = 3–6). * p < 0.05, ** p < 0.01, *** p < 0.001. α-actin ( ACTA2 ), smooth muscle myosin heavy chain (SM-MHC, MYH11 ), calponin ( CNN1 ), and synaptopodin-2 ( SYNPO2 ).

Article Snippet: Proteins were detected using commercially available primary antibodies against GATA6 (Cell Signaling #4253S, 1:1,000), α-actin (Sigma-Aldrich #A5228, 1:1,000), smooth muscle myosin heavy chain (abcam #ab53219, 1:1,000), calponin (abcam #ab46794, 1:1,000), SMAD1 (cell signaling, #9743, 1:1000), SMAD3 (cell signaling, SMAD 2/3 antibody, #3102, 1:1000), Hsp90 (BD Biosciences #610418, 1:10,000) at 4°C overnight.

Techniques: Cell Differentiation, Transduction, Expressing, Transfection

Journal: Frontiers in Physiology

Article Title: Transcription factor GATA6 promotes migration of human coronary artery smooth muscle cells in vitro

doi: 10.3389/fphys.2022.1054819

Figure Lengend Snippet: GATA6 promotes migration of human vascular smooth muscle cells. Human coronary artery smooth muscle cells were transduced with Ad-CMV-GATA6/Ad-CMV-null or transfected with GapmeRs against GATA6. Cell migration was assessed using a (A) wound-healing assay, (B,C) transwell migration assay (24 h). (D) Digital holographic cytometry was used to measure migration of human coronary artery smooth muscle cells after GapmerRs transfection to downregulate GATA6. Photos were taken at the indicated time points. Number of migrating cells were analyzed after 24 h. Migration speed and distance after 19 h. Data are presented as mean ± SEM ( n = 3–6). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Proteins were detected using commercially available primary antibodies against GATA6 (Cell Signaling #4253S, 1:1,000), α-actin (Sigma-Aldrich #A5228, 1:1,000), smooth muscle myosin heavy chain (abcam #ab53219, 1:1,000), calponin (abcam #ab46794, 1:1,000), SMAD1 (cell signaling, #9743, 1:1000), SMAD3 (cell signaling, SMAD 2/3 antibody, #3102, 1:1000), Hsp90 (BD Biosciences #610418, 1:10,000) at 4°C overnight.

Techniques: Migration, Transduction, Transfection, Wound Healing Assay, Transwell Migration Assay, Cytometry

Journal: Frontiers in Physiology

Article Title: Transcription factor GATA6 promotes migration of human coronary artery smooth muscle cells in vitro

doi: 10.3389/fphys.2022.1054819

Figure Lengend Snippet: Analysis of gene expression data from microarray of the cells overexpressing GATA6 and control. (A) Pie chart of pathway gene analysis of differentially expressed genes. Analysis was done using PANTHER (17.0 Release). The numbers represent fold enrichment. Only results with false discovery rate < 0.05 are presented. (B) Prediction of master regulators was done by QIAGEN Ingenuity Pathway Analysis software. A Network of master regulator was done by STRING database (version 11.5). The color shades represent the degree of activation/inhibition ranging from dark blue (most active) to dark red (most inhibited). The lines indicate the strength of data support such interaction. 17 master regulators were excluded from the network, since there was no evidence supporting any interaction with the other master regulators.

Article Snippet: Proteins were detected using commercially available primary antibodies against GATA6 (Cell Signaling #4253S, 1:1,000), α-actin (Sigma-Aldrich #A5228, 1:1,000), smooth muscle myosin heavy chain (abcam #ab53219, 1:1,000), calponin (abcam #ab46794, 1:1,000), SMAD1 (cell signaling, #9743, 1:1000), SMAD3 (cell signaling, SMAD 2/3 antibody, #3102, 1:1000), Hsp90 (BD Biosciences #610418, 1:10,000) at 4°C overnight.

Techniques: Gene Expression, Microarray, Control, Software, Activation Assay, Inhibition

Journal: Frontiers in Physiology

Article Title: Transcription factor GATA6 promotes migration of human coronary artery smooth muscle cells in vitro

doi: 10.3389/fphys.2022.1054819

Figure Lengend Snippet: Identification of genes regulated by GATA6 expression. A complete microarray screen of GATA6-sensitive gene transcription was performed on human coronary artery smooth muscle cells transduced with 100 MOI Ad-CMV-GATA6 or Ad-CMV-null (vehicle) as a control. Expression values for selected genes from the microarray from each sample ( n = 6) (A) . Validation of selected genes by qRT-PCR (B–D) and western blot (E) . Data are presented as mean ± SEM ( n = 6). ** p < 0.01, *** p < 0.001.

Article Snippet: Proteins were detected using commercially available primary antibodies against GATA6 (Cell Signaling #4253S, 1:1,000), α-actin (Sigma-Aldrich #A5228, 1:1,000), smooth muscle myosin heavy chain (abcam #ab53219, 1:1,000), calponin (abcam #ab46794, 1:1,000), SMAD1 (cell signaling, #9743, 1:1000), SMAD3 (cell signaling, SMAD 2/3 antibody, #3102, 1:1000), Hsp90 (BD Biosciences #610418, 1:10,000) at 4°C overnight.

Techniques: Expressing, Microarray, Transduction, Control, Biomarker Discovery, Quantitative RT-PCR, Western Blot

Journal: Frontiers in physiology

Article Title: Transcription factor GATA6 promotes migration of human coronary artery smooth muscle cells in vitro .

doi: 10.3389/fphys.2022.1054819

Figure Lengend Snippet: FIGURE 1 GATA6 induces vascular smooth muscle cell differentiation. Human coronary artery smooth muscle cells were transduced with Ad-CMV- GATA6/Ad-CMV-null (A,B). Expression of GATA6 and some smooth muscle markers are presented in (A) at mRNA level and (B) at protein level. (C) Relative mRNA expression of targets after transfection with GapmeRs against GATA6. Data are presented as mean ± SEM (n = 3–6). *p < 0.05, **p < 0.01, ***p < 0.001. α-actin (ACTA2), smooth muscle myosin heavy chain (SM-MHC, MYH11), calponin (CNN1), and synaptopodin-2 (SYNPO2).

Article Snippet: Proteins were detected using commercially available primary antibodies against GATA6 (Cell Signaling #4253S, 1:1,000), αactin (Sigma-Aldrich #A5228, 1:1,000), smooth muscle myosin heavy chain (abcam #ab53219, 1:1,000), calponin (abcam #ab46794, 1:1,000), SMAD1 (cell signaling, #9743, 1:1000), SMAD3 (cell signaling, SMAD 2/3 antibody, #3102, 1:1000), Hsp90 (BD Biosciences #610418, 1:10,000) at 4°C overnight.

Techniques: Cell Differentiation, Transduction, Expressing, Transfection

Journal: Frontiers in physiology

Article Title: Transcription factor GATA6 promotes migration of human coronary artery smooth muscle cells in vitro .

doi: 10.3389/fphys.2022.1054819

Figure Lengend Snippet: FIGURE 2 GATA6 promotes migration of human vascular smooth muscle cells. Human coronary artery smooth muscle cells were transduced with Ad- CMV-GATA6/Ad-CMV-null or transfected with GapmeRs against GATA6. Cell migration was assessed using a (A) wound-healing assay, (B,C) transwell migration assay (24 h). (D) Digital holographic cytometry was used to measure migration of human coronary artery smooth muscle cells after GapmerRs transfection to downregulate GATA6. Photos were taken at the indicated time points. Number of migrating cells were analyzed after 24 h. Migration speed and distance after 19 h. Data are presented as mean ± SEM (n = 3–6). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Proteins were detected using commercially available primary antibodies against GATA6 (Cell Signaling #4253S, 1:1,000), αactin (Sigma-Aldrich #A5228, 1:1,000), smooth muscle myosin heavy chain (abcam #ab53219, 1:1,000), calponin (abcam #ab46794, 1:1,000), SMAD1 (cell signaling, #9743, 1:1000), SMAD3 (cell signaling, SMAD 2/3 antibody, #3102, 1:1000), Hsp90 (BD Biosciences #610418, 1:10,000) at 4°C overnight.

Techniques: Migration, Transduction, Transfection, Wound Healing Assay, Transwell Migration Assay, Cytometry

Journal: Frontiers in physiology

Article Title: Transcription factor GATA6 promotes migration of human coronary artery smooth muscle cells in vitro .

doi: 10.3389/fphys.2022.1054819

Figure Lengend Snippet: FIGURE 3 Analysis of gene expression data from microarray of the cells overexpressing GATA6 and control. (A) Pie chart of pathway gene analysis of differentially expressed genes. Analysis was done using PANTHER (17.0 Release). The numbers represent fold enrichment. Only results with false discovery rate < 0.05 are presented. (B) Prediction of master regulators was done by QIAGEN Ingenuity Pathway Analysis software. A Network of master regulator was done by STRING database (version 11.5). The color shades represent the degree of activation/inhibition ranging from dark blue (most active) to dark red (most inhibited). The lines indicate the strength of data support such interaction. 17 master regulators were excluded from the network, since there was no evidence supporting any interaction with the other master regulators.

Article Snippet: Proteins were detected using commercially available primary antibodies against GATA6 (Cell Signaling #4253S, 1:1,000), αactin (Sigma-Aldrich #A5228, 1:1,000), smooth muscle myosin heavy chain (abcam #ab53219, 1:1,000), calponin (abcam #ab46794, 1:1,000), SMAD1 (cell signaling, #9743, 1:1000), SMAD3 (cell signaling, SMAD 2/3 antibody, #3102, 1:1000), Hsp90 (BD Biosciences #610418, 1:10,000) at 4°C overnight.

Techniques: Gene Expression, Microarray, Control, Software, Activation Assay, Inhibition

Journal: Frontiers in physiology

Article Title: Transcription factor GATA6 promotes migration of human coronary artery smooth muscle cells in vitro .

doi: 10.3389/fphys.2022.1054819

Figure Lengend Snippet: FIGURE 4 Identification of genes regulated by GATA6 expression. A complete microarray screen of GATA6-sensitive gene transcription was performed on human coronary artery smooth muscle cells transduced with 100 MOI Ad-CMV-GATA6 or Ad-CMV-null (vehicle) as a control. Expression values for selected genes from the microarray from each sample (n = 6) (A). Validation of selected genes by qRT-PCR (B–D) and western blot (E). Data are presented as mean ± SEM (n = 6). **p < 0.01, ***p < 0.001.

Article Snippet: Proteins were detected using commercially available primary antibodies against GATA6 (Cell Signaling #4253S, 1:1,000), αactin (Sigma-Aldrich #A5228, 1:1,000), smooth muscle myosin heavy chain (abcam #ab53219, 1:1,000), calponin (abcam #ab46794, 1:1,000), SMAD1 (cell signaling, #9743, 1:1000), SMAD3 (cell signaling, SMAD 2/3 antibody, #3102, 1:1000), Hsp90 (BD Biosciences #610418, 1:10,000) at 4°C overnight.

Techniques: Expressing, Microarray, Transduction, Control, Biomarker Discovery, Quantitative RT-PCR, Western Blot