Figure 6

ER-independent activities after AGR2 knockdown. T47 D cells were transfected with iNC or iAGR2, and after 24 hours, cells were treated in combination with 4-hydroxytamoxifen (4-OHT) or ICI 182,780 (ICI) at doses of 100 nM or 1 μM. The growth-inhibitory effects of combination treatments were assessed with Cell Titer Glo 96 hours after transfection (a), or the level of target modulation at 72 hours with Western blot analysis (b). (c) Modulation of p-Src was analyzed 72 hours after AGR2 knockdown. The kinetics of cyclin D1 and ER modulation were determined with Western blot, and densitometric values of cyclin D1 and ER were calculated and normalized to β-actin (Image J) and represented as the mean ± SD, n = 3. *P < 0.05; **P < 0.01; ***P < 0.001, antiestrogen and AGR2 knockdown combination versus AGR2 knockdown alone. Note: Some blots may be from different gels run with the same set of samples.