14022s pe anti human ptk7  (Cell Signaling Technology Inc)

Journal: Cell death & disease

Article Title: Recruitment of LEF1 by Pontin chromatin modifier amplifies TGFBR2 transcription and activates TGFβ/SMAD signalling during gliomagenesis.

doi: 10.1038/s41419-022-05265-y

Figure Lengend Snippet: Fig. 6 Pontin interacts with LEF1 to activate TGFBR2 gene transcription in GBM cells. a Venn plot showed the overlapping of Pontin interacting proteins with possible transcription factors regulating TGFBR2 gene expression. SF splicing factors, TF transcription factors. b Representative images showing that Pontin (red) colocalized with LEF1 (green) in U87MG and U251 cell nuclei. Scale bar, 10 μm. c Co-IP confirmation of the interaction between Pontin (flag-tagged) and endogenous LEF1 (upper) or HA-tagged LEF1 (bottom) in U87MG cells. d Effect of Pontin or LEF1 on the luciferase transcriptional response of the reporter driven by TGFBR2 promoter (−2000 to 0). e LEF1-binding sites predicted in the TGFBR2 promoter (left) and ChIP-qPCR analyses of the capacities for them to bind with LEF1 (right). Data in d, e are expressed as means ± SDs, n = 3.

Article Snippet: The primary antibodies were as follows: mouse anti-human Pontin (catalogue SAB4200194; Sigma‒Aldrich), mouse anti-human GAPDH (catalogue BM3876; Boster, China), goat anti-human TGFβRII (catalogue AF241-NA; Biotechne, USA), rabbit anti-human CTGF (catalogue A11067; Abclonal, China), rabbit anti-human SMAD2/SMAD3 (catalogue 5678; Cell Signaling Technology, USA), rabbit anti-human phospho-SMAD2/SMAD3 (catalogue 8828; Cell Signaling Technology), and rabbit anti-human LEF1 (catalogue 2230; Cell Signaling Technology).

Techniques: Gene Expression, Co-Immunoprecipitation Assay, Luciferase, Binding Assay, ChIP-qPCR

Journal: Cell death & disease

Article Title: Recruitment of LEF1 by Pontin chromatin modifier amplifies TGFBR2 transcription and activates TGFβ/SMAD signalling during gliomagenesis.

doi: 10.1038/s41419-022-05265-y

Figure Lengend Snippet: Fig. 6 Pontin interacts with LEF1 to activate TGFBR2 gene transcription in GBM cells. a Venn plot showed the overlapping of Pontin interacting proteins with possible transcription factors regulating TGFBR2 gene expression. SF splicing factors, TF transcription factors. b Representative images showing that Pontin (red) colocalized with LEF1 (green) in U87MG and U251 cell nuclei. Scale bar, 10 μm. c Co-IP confirmation of the interaction between Pontin (flag-tagged) and endogenous LEF1 (upper) or HA-tagged LEF1 (bottom) in U87MG cells. d Effect of Pontin or LEF1 on the luciferase transcriptional response of the reporter driven by TGFBR2 promoter (−2000 to 0). e LEF1-binding sites predicted in the TGFBR2 promoter (left) and ChIP-qPCR analyses of the capacities for them to bind with LEF1 (right). Data in d, e are expressed as means ± SDs, n = 3.

Article Snippet: Mouse anti-human Pontin antibody (catalogue SAB4200194; Sigma‒Aldrich) and rabbit anti-human LEF1 antibody (catalogue 2230; Cell Signaling Technology) were used as the primary antibodies.

Techniques: Gene Expression, Co-Immunoprecipitation Assay, Luciferase, Binding Assay, ChIP-qPCR