snai2 sirna  (Santa Cruz Biotechnology)

Journal: iScience

Article Title: MircroRNA-92b as a negative regulator of the TGF-β signaling by targeting the type I receptor

doi: 10.1016/j.isci.2023.108131

Figure Lengend Snippet: SNAI1 and SNAI2 inhibited miR-92b expression (A) The expression of miR-92b-3p in HK2 cells treated with TGF-β for 12 h, as detected by qPCR, n = 5. (B) Sequence logo of transcription factor SNAI1 and SNAI2. (C–E) Protein levels of SNAI1, SNAI2, and β-actin in HK2 cells treated with SNAI1 siRNA (siSNAI1), SNAI2 siRNA (siSNAI2), or control siRNA as indicated in the figure, as detected by western blotting; quantitative results are shown; n = 3. Expression of Col1a1 and miR-92b-3p as determined by qPCR; n = 5. (F) Schematic representation of the human THBS3-AS1 promoters with the putative E2-box promoter shown as a red box. (G) A series of truncated THBS3-AS1 promoters fused to the luciferase reporter gene was co-transfected into HEK293T cells under TGF-β stimulation together with the pcDNA3.1 Renilla plasmid (Ctrl). The relative luciferase activity was presented as folds of Ctrl (n = 3). (H) DNA fragments containing the flanking region of the E2-box on the THBS3-AS1 promoters were immunoprecipitated with anti-SNAI1 or anti-SNAI2 antibodies. Data are expressed as means ± SD; n = 6 per group; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Similar results from three independent experiments are shown.

Article Snippet: SNAI2 siRNA , Santa Cruz biotechnology , Cat# sc-38393.

Techniques: Expressing, Sequencing, Control, Western Blot, Luciferase, Transfection, Plasmid Preparation, Activity Assay, Immunoprecipitation

Journal: iScience

Article Title: MircroRNA-92b as a negative regulator of the TGF-β signaling by targeting the type I receptor

doi: 10.1016/j.isci.2023.108131

Figure Lengend Snippet:

Article Snippet: SNAI2 siRNA , Santa Cruz biotechnology , Cat# sc-38393.

Techniques: Recombinant, Control, Plasmid Preparation, Software, Modification, Staining, Extraction, Bicinchoninic Acid Protein Assay, Western Blot, cDNA Synthesis, SYBR Green Assay, Immunohistochemistry, Isolation

Journal: PLoS ONE

Article Title: The Transcription Factors Tbx18 and Wt1 Control the Epicardial Epithelial-Mesenchymal Transition through Bi-Directional Regulation of Slug in Murine Primary Epicardial Cells

doi: 10.1371/journal.pone.0057829

Figure Lengend Snippet: (A) Representative images of primary epicardial cells transfected with control siRNA (siControl) or siRNA directed against Tbx18 (siTbx18) or Wt1 (siWt1). Scale bar: 200 µm. (B) Immunostaining for ZO-1 or N-cadherin (green) and DAPI nuclear staining (blue) of primary epicardial cells transfected with siRNAs. Scale bars: 100 µm. (C) Percentage of cells categorized as “Enlarged” or “Cobblestone-like,” based on the cellular morphology of primary epicardial cells transfected with siRNAs. (D) Representative images of primary epicardial cells transfected with siRNAs at 0 and 14 hr after the scratch was made. Scale bar: 200 µm. (E) Quantification of migration distance, given as a ratio to the siControl (n = 4; * P <0.01 vs. siControl). (F) The relative mRNA expression of Tbx18 , Wt1 , Snail and Slug by real-time PCR analysis (n = 3; * P <0.05 vs. siControl). The results were normalized to Gapdh expression, and the relative expression level is given as a ratio to the siControl. (G) Western blot performed with antibodies against Tbx18, Wt1, adhesion molecules (E-cadherin and N-cadherin) and EMT regulators (Snail and Slug). β-actin and histone H3 were used as loading controls. The data are presented as the mean ± SD.

Article Snippet: The following siRNAs were used: Tbx18 siRNA (siTbx18; MSS233533, siTbx18-2; MSS233532, Invitrogen), Wt1 siRNA (siWt1; MSS212627, siWt1-2; MSS212628, Invitrogen), Slug siRNA (siSlug; MSS237944, Invitrogen) and control siRNA (siControl; SIC-001, Sigma-Aldrich).

Techniques: Transfection, Immunostaining, Staining, Migration, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: PLoS ONE

Article Title: The Transcription Factors Tbx18 and Wt1 Control the Epicardial Epithelial-Mesenchymal Transition through Bi-Directional Regulation of Slug in Murine Primary Epicardial Cells

doi: 10.1371/journal.pone.0057829

Figure Lengend Snippet: (A) Representative images of epicardial cells treated with TGFβ1 and transfected with control siRNA (siControl) or siRNA directed against Tbx18 (siTbx18), Slug (siSlug) or Wt1 (siWt1). (B) Immunostaining for ZO-1 (green) and DAPI nuclear staining (blue) of primary epicardial cells transfected with siRNAs. (C) Percentage of cells categorized as “Enlarged” or “Cobblestone-like,” based on cellular morphology. (D) The relative mRNA expression of Tbx18 , Wt1 , Snail and Slug by real-time PCR analysis in TGFβ1-treated epicardial cells transfected with siControl, siTbx18, siSlug or siWt1 (n = 3; * P <0.05 vs. siControl). The results were normalized to Gapdh expression, and the relative expression level is given as a ratio to the TGFβ1+siControl. (E) Western blot performed with antibodies against Tbx18, Wt1, Snail and Slug. β-actin and histone H3 were used as loading controls. The data are presented as the mean ± SD. Scale bars: 100 µm.

Article Snippet: The following siRNAs were used: Tbx18 siRNA (siTbx18; MSS233533, siTbx18-2; MSS233532, Invitrogen), Wt1 siRNA (siWt1; MSS212627, siWt1-2; MSS212628, Invitrogen), Slug siRNA (siSlug; MSS237944, Invitrogen) and control siRNA (siControl; SIC-001, Sigma-Aldrich).

Techniques: Transfection, Immunostaining, Staining, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: PLoS ONE

Article Title: The Transcription Factors Tbx18 and Wt1 Control the Epicardial Epithelial-Mesenchymal Transition through Bi-Directional Regulation of Slug in Murine Primary Epicardial Cells

doi: 10.1371/journal.pone.0057829

Figure Lengend Snippet: (A) Representative images of Wt1-knockdown epicardial cells co-transfected with siTbx18 or siSlug. Epicardial cells transfected with siControl or siWt1 were used as controls. (B) Immunostaining for ZO-1 (green) and DAPI nuclear staining (blue) of primary epicardial cells transfected with siRNA. (C) Percentage of cells categorized as “Enlarged” or “Cobblestone-like,” based on cellular morphology. (D) The relative mRNA expression of Tbx18 , Wt1 and Slug by real-time PCR in Wt1-knockdown epicardial cells co-transfected with siTbx18 or siSlug, in comparison to siControl-transfected epicardial cells (n = 3; * P <0.05 vs. siControl, † P <0.05 vs. siWt1). The results were normalized to Gapdh expression and the relative expression level is provided as a ratio to the siControl. The data are presented as the mean ± SD. Scale bars: 100 µm.

Article Snippet: The following siRNAs were used: Tbx18 siRNA (siTbx18; MSS233533, siTbx18-2; MSS233532, Invitrogen), Wt1 siRNA (siWt1; MSS212627, siWt1-2; MSS212628, Invitrogen), Slug siRNA (siSlug; MSS237944, Invitrogen) and control siRNA (siControl; SIC-001, Sigma-Aldrich).

Techniques: Transfection, Immunostaining, Staining, Expressing, Real-time Polymerase Chain Reaction