Fig. 6: MALAT1 and ID4 induce circ_0076611 expression in TNBC cells.

a RT-PCR analysis of circ_0076611 and of VEGFA linear transcripts in control (si-CTR) MDA-MB-468 cells and in cells depleted of ID4/MALAT1/mutant p53 expression, by transfection of siRNAs to ID4 and p53, as well as of ASO to MALAT1, has been performed using the primers indicated on the right. The identity of PCR products has been verified by Sanger sequencing and is indicated on the right. Sequences of siRNA and ASO are enclosed in Supplementary Table 1. b Expression of circ_0076611 (left) and MALAT1 (right), evaluated by RT-qPCR in the indicated breast cancer cell lines transfected with siRNAs or ASOs directed to MALAT1 (si-MALAT1#1, si-MALAT1#2, ASO-MALAT1) or controls (si-SCR, ASO-SCR). c–e Expression of circ_0076611 in MDA-MB-468 and M6 breast cancer cells silenced for ID4, by siRNA transfection (d) or by CRISPR/Cas9-mediated gene editing (d, e). f Western blot of ID4 protein in the experimental conditions analyzed in c–e. g Western blot analysis of the indicated circ_0076611 targets in control (CTR) MDA-MB-468 and in CRISPR/Cas9-mediated ID4-KO cells. Data are presented as mean plus standard deviation. *p ≤ 0.05, **p ≤ 0.005, **p ≤ 0.0o05 (paired, two-tailed Student’s t test). Results from at least three biological replicates are shown.