Fig. 2: Characterization of FGD5-AS1, EPB41L4A-AS1 and PVT1 as ERα-interacting lncRNAs. | Cell Death & Disease

Fig. 2: Characterization of FGD5-AS1, EPB41L4A-AS1 and PVT1 as ERα-interacting lncRNAs.

From: LncRNA PVT1 links estrogen receptor alpha and the polycomb repressive complex 2 in suppression of pro-apoptotic genes in hormone-responsive breast cancer

Fig. 2

A Box Plots showing lncRNAs expression levels (-log2(TPM + 1)) comparing tumor (red) and normal (gray) tissues from TCGA data by using GEPIA2 tool. The number of the tissues considered in the analysis is listed below each comparison. Asterisk indicates statistically significant differences using One-Way ANOVA (*p < 0.05). B Bar plots showing the expression levels of the selected lncRNAs in ERα-positive (MCF-7, T-47D and ZR-75-1), ERα-negative (Hs-578T, MDA-MB-231) BC cell lines an in the mammary epithelial cell line MCF-10A. RT-qPCR results shown are the mean ± SD of triplicate values using as housekeeping internal control RPLP0. C Bar plots showing RT-qPCR results of fractionated (cytosol and nucleus) lncRNAs distribution relative to total RNAs. MALAT1 and GAPDH were employed as housekeeping controls for nucleus and cytosol respectively. The results showed are the mean ± SD of triplicate values. Asterisks indicate statistically significant differences using unpaired t-test (**p < 0.01, ***p < 0.005 and ****p < 0.001). D Bar plots showing the expression levels (measured with RT-qPCR) of TFF1 mRNA (left) and of the three lncRNAs (right) following 24 h of mitogenic E2 stimulation in hormone-deprived MCF-7. RPLP0 was used as housekeeping. The results showed are the mean ± SD of three independent experiments. Asterisks indicate statistically significant differences using unpaired t-test (**p < 0.01 and ****p < 0.001). E Bar plots showing RT-qPCR results of relative fold expression levels of the indicated transcripts following ESR1 knock-down. ESR1 and TFF1 mRNA levels were analyzed as experimental controls. Data were analyzed with respect to the scramble (Silencer Select Negative Control: CTRL). The results showed are the mean ± SD of three independent experiments. Asterisks indicate statistically significant differences using unpaired t-test (****p < 0.001). F Chromatin associated RNA immunoprecipitation (CARIP) coupled to RT-qPCR showing the enrichment of ERα-associated lncRNAs on MCF-7 BC cell chromatin. IgG was used as negative control. lncRNA MALAT1 was used as positive experimental control. The results showed are the mean ± SD of triplicate values. Asterisks indicate statistically significant differences using unpaired t-test (*p < 0.05, **p < 0.01 and ****p < 0.001).

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