Fig. 3

Loss of XBP1 attenuates induction of estrogen-responsive genes and sensitizes MCF7 cells to anti-estrogens. A After synchronization, cells were either treated with (Vehicle) DMSO or (10 nM E2) estrogen. Line graphs show change in O.D at 490 nm after E2 treatment. Data shown is mean ± S.D (n = 6). *p < 0.05, two tailed unpaired t-test. B Cells were treated with cycloheximide (CHX, 100 μg/ml) and expression of ERα and β-Actin was determined by western blotting. Representative western blot from three independent experiments is shown. C MCF7 control and XBP1 KO cells were treated with either vehicle (DMSO) or (10 nM) E2 for the required time points. Relative expression of indicated genes was analysed by qRT-PCR (n = 3), RPLP0 was used to normalize the gene expression. Data presented as mean ± SD. p-values were determined using two-tailed unpaired t-test, *p < 0.05, **p < 0.01, ns- not significant. (D) Cells were treated with (10 µM) tamoxifen and (10 µM) fulvestrant for indicated time points. Line graphs show change in O.D at 490 nm. Data presented is mean ± S.D of five independent experiments. *p < 0.05, two-tailed unpaired t-test comparing respective time points