Journal: Frontiers in Oncology
Article Title: Activin A Signaling Regulates IL13Rα2 Expression to Promote Breast Cancer Metastasis
doi: 10.3389/fonc.2019.00032
Figure Lengend Snippet: INHBA is correlated with IL13Rα2 expression in breast cancer cells. (A) MII cells were cultured in DMEMF/12 medium containing 0.2% horse serum (HS) overnight and then treated with 10 ng/ml Activin A for different time points. Western blotting was performed using whole cell lysates to assess phosphorylation of Smad2 (Ser465/467), Smad3 (Ser423/425), Akt (Ser 473), and Erk1/2 (Thr202/Tyr204). Detection of total Smad2, Smad3, Akt, and Erk1/2 was used as loading controls. Protein expression was quantified using ImageJ software. (B) MII cells were mock-treated (DMSO) or treated 1 μM EW-7197 or SB-505124, in the presence or absence of 10 ng/ml Activin A for 1 h. Western blotting was performed using whole cell lysates to assess the phosphorylation status of Smad2 and Smad3. Total Smad2, Smad3, and β-actin were detected as loading controls. (C) MIV cells were treated with 1 μM EW-7197 or 1μM SB-505124 for 24 h. Real time qPCR was used to measure the mRNA expression of IL13Rα2. Asterisk ( * ) indicates a statistically significant difference between Activin A-treated and mock-treated cells ( n = 6; p < 0.05). (D) MIV cells were stably transduced with lentiviral vectors expressing shSCR or shSmad2. Real time qPCR was used to measure the mRNA expression of IL13Rα2. Asterisk ( * ) indicates a statistically significant difference between MIV-shSCR and MIV-shSmad2 ( n = 3; p < 0.05). (E) Western blotting showing the protein expression of Smad2 and IL13Rα2 in MIV-shSCR and MIV-shSmad2 cells. Antibody against β-tubulin was used as a loading control. Protein expression was quantified using ImageJ software.
Article Snippet: To study the effects of small molecule inhibitors on Activin A-induced gene expression, MII cells were serum starved in 0.2% HS-containing DMEM/F12 overnight and then treated with 1 μM EW-7197 (TargetMol), 1 μM SB-505124 (TargetMol) or 10 ng/ml Activin A alone or in combination (as indicated in respective figures).
Techniques: Expressing, Cell Culture, Western Blot, Software, Stable Transfection, Transduction
TargetMol manufactures this product 
