Journal: Oncotarget

Article Title: Crosstalk between integrin αvβ3 and ERα contributes to thyroid hormone-induced proliferation of ovarian cancer cells

doi: 10.18632/oncotarget.10757

Figure Lengend Snippet: ( A ) mRNA levels of ERα , TRβ1 , ITG αv and ITG β3 from MCF-7, SKOV-3 and OVCAR-3 were measured by quantitative real-time PCR and normalized with 18Sr RNA. Results were expressed as folds of increase. ( B ) SKOV-3 cells were treated in the presence or absence of indicated peptides with T 3 or T 4 for 3 days, cells in 96 wells were subjected to the MTT assay. ( C ) scramble or shRNA of integrin αv or β3 were transiently transfected to SKOV-3 cells and treated with thyroid hormone (T 4 ) for 30 min, and whole cell lysates were harvested for western blot analysis. Indicated antibodies were used. Compared to sc: p < 0.05 :* p < 0.01 :** p < 0.001 :*** ; Compared to sc treated with T 4 : p < 0.05: † p < 0.01: †† p < 0.001: †††

Article Snippet: Membranes were blocked with 5% milk in Tris-buffered saline containing 0.1% Tween, and then incubated with selected antibodies overnight: PCNA (GTX100539, GeneTex Inc. CA, USA), Lamin B (GTX103292, GeneTex Inc.), integrin αv (SC9969, Santa Cruz Inc., Santa Cruz, CA, USA ), integrin β3 (SC6627, Santa Cruz), phospho-ERK1/2 (4377S, Cell Signaling Technology, Danvers, MA, USA), total-ERα (8644S, Cell Signaling Technology), ERα-pS118(2511p, Cell Signaling Technology), ERα-pS167 (5587p, Cell Signaling Technology).

Techniques: Real-time Polymerase Chain Reaction, MTT Assay, shRNA, Transfection, Western Blot

Journal: Oncotarget

Article Title: Crosstalk between integrin αvβ3 and ERα contributes to thyroid hormone-induced proliferation of ovarian cancer cells

doi: 10.18632/oncotarget.10757

Figure Lengend Snippet: SKOV- 3 cells were treated with ( A ) thyroid hormone, ( B ) estrogen, for different periods of time as indicated. ( C ) SKOV-3 cells treated with thyroid hormone or estrogen for 10 min in the absence or presence of ICI were fixed and stained with anti-integrin αv and phospho-ERα (S167) antibodies, and subsequently with fluorescent secondary antibodies. Nuclear punctate of phosphorylated ERα-induced by T 4 (shown in green) were increased with time and co-localized with integrin αvβ3 (shown in red) to yield a yellow color. Nuclei were stained with DAPI and showed as blue. The phosphorylation of ERα and nuclear translocation of integrin αv were inhibited by ICI. Quantitative fluorescence intensities are shown as average intensity per cell. p < 0.05: * p < 0.01: **

Article Snippet: Membranes were blocked with 5% milk in Tris-buffered saline containing 0.1% Tween, and then incubated with selected antibodies overnight: PCNA (GTX100539, GeneTex Inc. CA, USA), Lamin B (GTX103292, GeneTex Inc.), integrin αv (SC9969, Santa Cruz Inc., Santa Cruz, CA, USA ), integrin β3 (SC6627, Santa Cruz), phospho-ERK1/2 (4377S, Cell Signaling Technology, Danvers, MA, USA), total-ERα (8644S, Cell Signaling Technology), ERα-pS118(2511p, Cell Signaling Technology), ERα-pS167 (5587p, Cell Signaling Technology).

Techniques: Staining, Phospho-proteomics, Translocation Assay, Fluorescence

Journal: Oncotarget

Article Title: Crosstalk between integrin αvβ3 and ERα contributes to thyroid hormone-induced proliferation of ovarian cancer cells

doi: 10.18632/oncotarget.10757

Figure Lengend Snippet: SKOV-3 were pre-treated in the presence or absence of ICI for 30 min prior to another 30 min of T 4 treatment and harvested for ChIP. Total cell lysate was immunoprecipitated with anti-integrin αv and pulled-down DNA was measured with qPCR. Compared to control: p < 0.01: **; Compared to T 4 alone: p < 0.001: †††

Article Snippet: Membranes were blocked with 5% milk in Tris-buffered saline containing 0.1% Tween, and then incubated with selected antibodies overnight: PCNA (GTX100539, GeneTex Inc. CA, USA), Lamin B (GTX103292, GeneTex Inc.), integrin αv (SC9969, Santa Cruz Inc., Santa Cruz, CA, USA ), integrin β3 (SC6627, Santa Cruz), phospho-ERK1/2 (4377S, Cell Signaling Technology, Danvers, MA, USA), total-ERα (8644S, Cell Signaling Technology), ERα-pS118(2511p, Cell Signaling Technology), ERα-pS167 (5587p, Cell Signaling Technology).

Techniques: Immunoprecipitation, Control