Fig 2: SNHG1 inhibited ox-LDL-induced cell injury and inflammatory response via up-regulating GNAI2 and PCBP1. (A, B) CCK-8 and MTT assay determined cell viability of indicated ox-LDL-induced cells. (C, G). Cell apoptosis rate (C), caspase-3/8/9 activity (D), LDH release and protein expression (E), IL-6 secretion and protein expression (F), IL-1ß secretion and protein expression (G) in ox-LDL-induced HUVECs in indicated groups. Four groups in the experiments: pcDNA3.1-NC, pcDNA3.1-SNHG1 (**P < 0.01 versus the 1st group), pcDNA3.1-SNHG1+sh-PCBP1 (*P < 0.05 versus the 2nd group), pcDNA3.1-SNHG1+sh-GNAI2/PCBP1 (**P < 0.01 versus the 2nd group). One-way ANOVA followed by Tukey post hoc test. Data obtained from more than three repeated experiments were shown as mean ± SD. * indicated P < 0.05 and ** indicated P < 0.01 meant data were statistically significant.

Fig 3: The Targeted Inhibition of PCBP1 and FUS Could Reduce the Malignant Progression of CML Cells(A) K562 cells and CML patient bone marrow mononuclear cells transfected with SCR, PCBP1 siRNA, and FUS siRNA were treated with different concentrations of imatinib (0–0.25 µM) for 48 h, and the viability of cells was determined by MTT assay (**p < 0.01, *p < 0.05, PCPB1-siRNA and FUS-siRNA transfected groups versus K562 group and SCR transfected group). (B) Inhibition of PCPB1 and FUS reduces colony formation of K562 cells. A total of 1,000 K562 cells transfected with SCR, PCBP1, and FUS were mixed with RPMI-1640 medium containing 0.9% methylcellulose solution and 20% FBS and seeded into 24-well plates. Colony numbers were counted after 1 week. Histogram and statistics indicating the relative number of colonies per 1,000 plated cells are shown. Statistical significance was assessed by one-way ANOVA (**p < 0.01). (C) K562 cells were co-transfected with H19 lentivirus and FUS plasmid, and the viability of cells was determined by MTT assay. (D) K562 cells were co-transfected with H19 lentivirus and PCBP1 plasmid, and the viability of cells was determined by MTT assay.

Fig 4: lncRNA H19 Could Target PCBP1 and FUS Protein and Could Target miR-19a-3p and miR-106b-5p in CML(A) PCBP1 and FUS proteins have been found in the mass spectra results of long non-coding RNA H19 pull-down products. catRAPID and starBase v. 2.0 predicted that PCBP1 and FUS are the interactive proteins of the long non-coding RNA H19. (B) The products of RNA pull-down were detected by immunoblotting using rabbit anti-PCBP1 antibody and using rabbit anti-FUS antibody. (C) lncRNA H19 targets were identified by combining starBase v. 2.0 prediction and RNA-seq results of lncRNA H19 pull-down products. The hsa-miR-19a-3p, hsa-miR-106b-5p, hsa-miR-148a-3p, hsa-miR-148b-3p, and hsa-miR-454-3p identified by both methods were considered as candidate targets of lncRNA H19. (D) The identification of miR-19a-3p and miR-106b-5p by real-time PCR.

Fig 5: E2 rescued primary cultured hippocampal neurons from erastin-induced ferroptosis. (A) Cell viability of primary cultured hippocampal neurons treated with or without erastin (20 µM), E2 (30 µM) and Lip-1 (10 µM) for 12 h (n = 3). (B) Relative RNA level of DHODH (n = 3). (C) Representative images of western blots and quantitative analysis showing the expression levels of DHODH, GPX4, TfR, ferritin, FPN1, PCBP1 and GAPDH in primary cultured hippocampal neurons (n = 3). (D) and (F) Representative images of BODIPY C11 581/591 staining (scale bar = 50 µm) and quantitative analysis of lipid peroxidation in hippocampal neurons by the green/red fluorescence ratio. The cell nuclei were stained with DAPI (blue). (E) and (G) Representative images of MitoSOX staining (scale bar = 50 µm) and quantitative analysis of the red/blue fluorescence ratio. (H) and (I) MMP changes were measured by FCM analysis of JC-1 using annexin-V fluorescein isothiocyanate (FITC)/propidium iodide (PI). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)