ttp22  (MedChemExpress)

Journal: eLife

Article Title: Differentiation of mouse fetal lung alveolar progenitors in serum-free organotypic cultures

doi: 10.7554/elife.65811

Figure Lengend Snippet: Figure 4. Compounds promoting AT1 cell differentiation lead to concomitant loss of AT2 marker gene expression and WNT-dependent transcription. (A) IC261, CX-4945, and DMAT treatments (48 h) lead to downregulation of AT2 cell markers. TTP22 and Emodin (also CK2 inhibitors) do not lead to the same phenotype, possibly due to differences in potency/target affinity. Expression levels relative to DMSO control. Three biological replicates (consisting of 2 technical replicates each) were analyzed per compound. (B) Casein Kinase inhibition by IC261, CX-4945, and DMAT (6 h) leads to reduced Axin2 relative expression. Treatment with TTP22 or Emodin does not significantly affect Axin2 levels. More than three biological replicates were analyzed. (C) CK inhibition reduced WNT/β-catenin-dependent transcription in HEK293T epithelial cells (SuperTOPFlash-based luciferase assay). (B) Mean values are displayed; error bars represent S.D.; p-values from one-way ANOVA, Tukey’s multiple comparisons test. (C) Mean values displayed; error bars represent S.D.; p-values from one-way ANOVA, Tukey’s multiple comparisons test; displayed p-values refer to comparisons with WNT3A- treated condition.

Article Snippet: or resource Designation Source or reference Identifiers Additional information Chemical compound, drug Emodin Tocris Tocris:3811 (10 μM) Chemical compound, drug TTP22 Tocris Tocris:4432 (10 μM) Chemical compound, drug IC261 Sigma Sigma:I0658 (10 μM) Chemical compound, drug CX- 4945 Enzo Life Sciences Enzo Life Sciences: ENZ- CHM151 (10 μM) Chemical compound, drug DMAT Sigma Sigma:SML2044 (10 μM) Chemical compound, drug IWR- 1 Sigma Sigma:I0161 (10 μM) Commercial assay or kit Dual- Luciferase Reporter Assay System Promega Promega:E1910 Commercial assay or kit NucleoSpin RNA kit Macherey- Nagel MachereyNagel:740955.50 Commercial assay or kit Superscript III Reverse Transcriptase system Invitrogen Thermo Fisher Scientific:18080093 Commercial assay or kit DyNAmo ColorFlash SYBR green qPCR kit Thermo Scientific Thermo Fisher Scientific:F416XL Sequencebased reagent qPCR This paper Supplementary file 1 Software, algorithm Fiji/ImageJ 1.53 c Schindelin et al., 2012; doi:10.1038/nmeth.2019 RRID:SCR_002285 Software, algorithm GraphPad Prism 8 GraphPad RRID:SCR_002798 Continued

Techniques: Cell Differentiation, Marker, Gene Expression, Expressing, Control, Inhibition, Luciferase

Journal: eLife

Article Title: Differentiation of mouse fetal lung alveolar progenitors in serum-free organotypic cultures

doi: 10.7554/eLife.65811

Figure Lengend Snippet: ( A ) IC261, CX-4945, and DMAT treatments (48 h) lead to downregulation of AT2 cell markers. TTP22 and Emodin (also CK2 inhibitors) do not lead to the same phenotype, possibly due to differences in potency/target affinity. Expression levels relative to DMSO control. Three biological replicates (consisting of 2 technical replicates each) were analyzed per compound. ( B ) Casein Kinase inhibition by IC261, CX-4945, and DMAT (6 h) leads to reduced Axin2 relative expression. Treatment with TTP22 or Emodin does not significantly affect Axin2 levels. More than three biological replicates were analyzed. ( C ) CK inhibition reduced WNT/β-catenin-dependent transcription in HEK293T epithelial cells (SuperTOPFlash-based luciferase assay). ( B ) Mean values are displayed; error bars represent S.D.; p - values from one-way ANOVA, Tukey’s multiple comparisons test. ( C ) Mean values displayed; error bars represent S.D.; p-values from one-way ANOVA, Tukey’s multiple comparisons test; displayed p-values refer to comparisons with WNT3A-treated condition. Figure 4—source data 1. Normalized expression values of AT2 cell markers, CK inhibitors vs. DMSO control. Normalized Axin2 expression values, CK inhibitors vs. DMSO control and related statistics. Raw luciferase data. Normalized luciferase values and related statistics.

Article Snippet: Chemical compound, drug , TTP22 , Tocris , Tocris:4432 , (10 μM).

Techniques: Expressing, Control, Inhibition, Luciferase

Journal: eLife

Article Title: Differentiation of mouse fetal lung alveolar progenitors in serum-free organotypic cultures

doi: 10.7554/eLife.65811

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , TTP22 , Tocris , Tocris:4432 , (10 μM).

Techniques: Recombinant, Transfection, Construct, Plasmid Preparation, Mutagenesis, Reporter Assay, Reverse Transcription, SYBR Green Assay, Sequencing, Software