MSN and NONO were amplified from the reverse-transcribed cDNA from MDA-MB-231 cell line and cloned into pSIN-FLAG (puromycin or blasticidin resistant) vector (Addgene, USA), and the authenticity was verified by sequencing, respectively. Primer design was used to introduce site mutation to construct MSN-overexpressing plasmids with different phosphorylation patterns. Plasmid constructs expressing short hairpin RNAs (shRNAs) were designed by Sigma-Aldrich; the sense sequence was designed as follows in Table 2.

A highly efficient lentiviral system was used to generate MSN- or NONO-overexpressing plasmid DNA and the shRNA plasmid DNA. The cell lines were infected with the lentiviruses, and the stable cell lines were established. The lentiviral transfection efficiency was more than 90% in all cell lines.

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