Journal: PLoS ONE

Article Title: n-Butyl Benzyl Phthalate Promotes Breast Cancer Progression by Inducing Expression of Lymphoid Enhancer Factor 1

doi: 10.1371/journal.pone.0042750

Figure Lengend Snippet: The expression of CRITPO, FGF, PKA, EDNRA, LMO1, VEGF-D, LEF-1, PDGFR, PDGFRA, ADCV, and TCF (LEF) was examined in R2d and MCF-7 cells treated with 1 µM BBP using qRT-PCR. The data represent the mean ± SE of three experiments.

Article Snippet: The following siRNAs were used: control siRNA (sense: 5′-GAU CAU ACG UGC GAU CAG A-3′ , antisense: 5′-UCU GAU CGC ACG UAU GAU C-3′ ); LEF-1 siRNA-1 (GenBank SASI_00151593; sense: 5′-GAA ACU ACA GGA AUC UGCA-3′ , antisense: 5′-UGC AGA UUC CUG UAG UUUC-3′ ); LEF-1 siRNA-2 (GenBank SASI_00349169; sense: 5′-CAU CAG AUG UCA ACU CCA A-3′ , antisense: 5′-UUG GAG UUG ACA UCU GAUG-3′ ).The following antibodies used for immunoblotting were purchased: anti-LEF-1(Origene Technologies, Inc., Rockville, MD) and anti-β-actin (Sigma)

Techniques: Expressing, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: n-Butyl Benzyl Phthalate Promotes Breast Cancer Progression by Inducing Expression of Lymphoid Enhancer Factor 1

doi: 10.1371/journal.pone.0042750

Figure Lengend Snippet: (A) One identified gene, LEF-1, was found to be in the proliferation pathway group, the EMT pathway group, and the angiogenesis pathway group. (B, C) Silencing of LEF-1 was performed by transfecting LEF-1 siRNA-1 (5 nM) or LEF-1 siRNA-2 (5 nM), and LEF-1 expression was detected with qRT-PCR, western blot and compared to cells transfected with control siRNA. (D, E) Breast cancer cells were pretransfected with control siRNA or siRNA for LEF-1 (siRNA-1/2) and then treated with BBP for 24 h. Cell viability was analyzed with the XXT assay in R2d (D) and MCF-7 (E) cells. Cells treated as in panels D and E were assessed with the invasion assay (F, G) and wound healing assay (H, I). The horizontal dashed line in panels H and I indicates the width of the wound. The data represent the mean ± SE of three experiments. Each asterisk denotes a significant difference compared to the control (P<0.05; one-tailed Student's t-test).

Article Snippet: The following siRNAs were used: control siRNA (sense: 5′-GAU CAU ACG UGC GAU CAG A-3′ , antisense: 5′-UCU GAU CGC ACG UAU GAU C-3′ ); LEF-1 siRNA-1 (GenBank SASI_00151593; sense: 5′-GAA ACU ACA GGA AUC UGCA-3′ , antisense: 5′-UGC AGA UUC CUG UAG UUUC-3′ ); LEF-1 siRNA-2 (GenBank SASI_00349169; sense: 5′-CAU CAG AUG UCA ACU CCA A-3′ , antisense: 5′-UUG GAG UUG ACA UCU GAUG-3′ ).The following antibodies used for immunoblotting were purchased: anti-LEF-1(Origene Technologies, Inc., Rockville, MD) and anti-β-actin (Sigma)

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Invasion Assay, Wound Healing Assay, One-tailed Test

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