Mean ± SEM, *p<0.05 and **p<0.01.įigure S4: p53 is a downstream mediator of MALAT1. (Bd) qRT-PCR analyses reveal that transiently expressed MALAT1 rescues the expression of cell cycle genes ( Mybl2, Ccna2, CenpE) in MALAT1-depleted HDFs. Note that several of the upregulated genes are part of the p53-signaling pathway ( Cdkn1a, Gadd45a, Gadd45b, Il-6, Il-8, Mdm2, Tp53inp1). (Bc) The relative expression of indicated genes is determined by qPCR using RNA from control (scr) and MALAT1-depleted (AS1 & AS2) WI-38 cells. Note that the p53-signaling pathway is activated in MALAT1-depleted lung fibroblasts. S2B: (Ba) Top significant biofunctions and (Bb) canonical pathways of the protein-coding genes that are upregulated in MALAT1-depleted fibroblasts. Note that MALAT1-depleted cells do not show changes in PCNA mRNA levels. (Ae) The relative expression of PCNA is determined by qRT-PCR (with 3 independent primer pairs) using RNA from control (scr) and MALAT1-depleted (AS1 & AS2) WI-38 cells. Note that MALAT1 depletion using double-stranded siRNA oligos also result in reduced expression of cell-cycle genes. (Ad) Changes in relative expression of indicated genes determined by qRT-PCR using total RNA from control (using control siRNA) and MALAT1-depleted (using MALAT1-specific siRNA) WI-38 cells. Mean ± SEM, *p<0.05, **p<0.01 and ***p<0.001.įigure S2: S2A (Aa–c) The relative expression of indicated genes determined by qRT-PCR from total RNA isolated from control (scr) and MALAT1-depleted (AS1 & AS2) WI-38 cells. (F) The relative expression of indicated genes is determined by qRT-PCR analyses from total RNA of control and MALAT1-depleted (using siRNA oligos) WI-38 cells. (E) Bright field low magnification images of control and MALAT1-depleted HDFs show changes in cellular morphology in MALAT1-depleted cells. (D) Flow cytometry analyses of control (control-si) and MALAT1-depleted (using MALAT1-si1, -si2 & -si3 double stranded siRNA oligos) WI-38 cells. (C) Flow cytometry analyses of control (scr-oligo) and MALAT1-depleted (MALAT1-AS1 & AS2 antisense oligonucleotides) WI-38 cells. (B) Expression of SRSF1 is determined by immunoblotting using extracts from cell cycle-synchronized U2OS cells. Note that the MALAT1 levels increased during G1/S phase (G1/S is determined by the increased levels of cyclin E). (A) MALAT1 and Cyclin E levels in 10 and 15 hr post release serum-starved WI-38 cells are analyzed by qRT-PCR.
Figure S1: MALAT1 levels are cell cycle regulated and depletion of MALAT1 results in proliferation defects.