Pglv5 h1 gfp puro vector

Lentivirus packaging cells were transfected with PGLV3-h1-GFP-puro vector (GenePharma, Shanghai, China) or pGLV5-h1-GFP-puro vector (GenePharma, Shanghai, China) containing either the RNPC1a knockdown (shRNPC1a) or RNPC1a overexpression (RNPC1a), and a scrambled sequence (SCR) or a negative control sequence (NC), respectively, following the manufacturer’s instructions. Three shRNA plasmids (sh1, sh2, sh3) were constructed against different RNPC1a targets, including a scrambled sequence as a negative control (Additional file 1: Table S1). All plasmids were verified by sequencing (GenePharma, Shanghai, China). Cells were plated in 6 wells dishes at 30% confluence and infected with the retroviruses. Meanwhile, polybrene (5 μg/ml) was added with the retroviruses to enhance the target cells infection efficiency. Stable pooled populations of breast cancer cells were generated by selection using puromycin (2 μg/ml) for 2 weeks. For knockdown, one construct (sh3), with ≥85% knockdown efficiency, was used for further studies (Additional file 1: Figure S1).

Lentivirus constructed cells were transfected with PGLV3-h1-GFP-puro vector (GenePharma, China) or pGLV5-h1-GFP-puro vector (GenePharma, China) containing either RBMS2 knockdown (shRBMS2) or RBMS2 overexpression (RBMS2), and a scrambled sequence (SCR) or a negative control sequence (NC), respectively, following the manufacturer’s instructions. All plasmids were verified by sequencing (GenePharma, China). Cells were plated in 6 wells dishes at 30% confluence and infected with the retroviruses. Meanwhile, polybrene (5 μg/ml) was added with the retroviruses to enhance the target cells infection efficiency. Stable pooled populations of breast cancer cells were generated by selection using puromycin (2 μg/ml) for 2 weeks.
MCF-7 and SUM-1315 RBMS2 overexpression (RBMS2) and the control (NC) cells were seeded in 6-well plates overnight and then transfected with P21-siRNA (GenePharma, China) and the nonspecific siRNA control, using Lipofectamine® 3000 transfection agent (Invitrogen, USA). The sequences of the siRNAs are as follows:

sense 5’-CCUCUGGCAUUAGAAUUAUTT-3′ and

antisense 5’-AUAAUUCUAAUGCCAGAGGTT-3’