Fig. 3.

Signaling alterations in the engineered T47D cells following endocrine treatment in vitro and in vivo. a Signaling alterations in the engineered T47D cells following endocrine treatment. Western blot analysis of the engineered T47D cells expressing ESR1–CCDC170 fusion variants or wtCCDC170 cultured in normal medium (RPMI 1640 with phenol and 10% FBS), estrogen-deprived (ED) medium (phenol red-free RPMI 1640 with 5% CSS), or ED medium plus 4-OH tamoxifen (0.5 μM) for 6 days. b The effect of ER depletion on the viability of engineered T47D cells expressing ESR1-CCDC170 variants or vector control as shown by clonogenic assays. Intensities of colonies in each well was normalized to the respective YFP group. The representative plate images are shown on the top. Upper panel, western blots verifying the knockdown efficiencies of siERα. *P < 0.05, **P < 0.01, ***P < 0.001 (Student’s t test). c Western blot analysis of protein extracts from the engineered T47D xenograft tumor tissues treated with tamoxifen. Densitometric results of western blots are shown in the figure. *P < 0.05,**P < 0.01 (Student’s t test)