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Fig. 2 | BMC Cancer

Fig. 2

From: Loss of REST in breast cancer promotes tumor progression through estrogen sensitization, MMP24 and CEMIP overexpression

Fig. 2

Loss of REST leads to activation of ER⍺ signaling. A Downregulation of REST upon siRNA transfection. MCF-7 cells were transfected with control siRNA or siRNA directed to REST. Twenty-four hours after transfection, cells were treated with vehicle (PBS), estradiol (E2), progesterone (P4) or both. RNA-sequencing was performed with technical replicates (n = 2). Forty-eight hours after transfection REST protein levels were assayed by immunoblotting. Blot has been cropped for clarity. B-actin was used as a loading control. B REST mRNA level in MCF-7 cells upon REST siRNA transfection in vehicle treated cells (n = 3) (Representative image for siREST in all treated groups). C Principal component analysis (PCA) plot indicates distinct gene expression patterns based on treatment. Gene expression induced by REST knock down is asserted along PCA-2. Estradiol induced gene expression is asserted along PCA-3. Progesterone induced gene expression is asserted along PCA-4 (not in figure). D IPA identified REST downstream target genes in a network that are upregulated upon REST siRNA transfection. E Estrogen receptor signaling was shown to be upregulated in the REST siRNA, vehicle treated cells compared to the control. Upregulated genes shown in red, downregulated genes shown in green, predicted activation shown in orange, and predicted inhibition shown in blue. Error bars indicate ± SD, Student’s T-test was performed, *P < 0.05

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