Fig. 4

EGFR and KNG1 expression in control and GnRH agonist treated LN229 cells using Western blot analysis. The densitometric analysis of the Western blot shows a significant reduction in EGFR (2.2 fold) (A) and overexpression of KNG1 (1.5 fold) (B) after GnRH agonist treatment in comparison to untreated cells. For Western blot analysis, a total of 15 μg protein from LN229 cell lysate (Control and GnRH agonist treated group) was resolved by SDS-PAGE and electrotransferred onto PVDF membrane. The membrane was incubated with primary antibody against EGFR (dilution 1:2000) and KNG1 (dilution 1:5000) followed by incubation with anti-rabbit secondary antibody (dilution 1:30,000). The blots were developed using ECL reagent, image was acquired using Chemidoc MP (Bio-Rad) followed by densitometric analysis. The bar diagram shows the expression level of EGFR (C) and KNG1 (D) in control and agonist treated cell lysate. Error bars represent the standard deviation of mean. Full-length blot images are presented in Supplementary Fig. 1C and D