Fig 1: Inducible gene modulation in murine melanoma cell lines. (a) Schematic depicting the generation of stable cell lines expressing inducible Cas9 and sgRNA targeting GFP or Mafg. Mouse melanoma cell lines were infected with a lentiviral construct expressing Dox-inducible Cas9 and selected with blasticidin. Subsequently, TRE-Cas9‒harboring cells were infected with GFP-U6-sgGFP or GFP-U6-sgMafg1 and selected with hygromycin, and polyclonal populations were used for experiments. (b) TRE-Cas9‒harboring mouse melanoma cell lines were infected with a GFP-U6-sgGFP reporter construct. Western blot showed that Dox treatment resulted in Cas9 (Flag) expression and a decrease in GFP levels. (c) TRE-Cas9‒harboring M10M3 and M167M1 cells were infected with GFP-U6-sgGFP or GFP-U6-sgMafg1. Cells were then treated with Dox, and Flag-Cas9, GFP, and MAFG expressions were analyzed by western blot. Cas9 was expressed on Dox treatment, GFP was decreased in sgGFP-expressing cells, and MAFG was decreased in sgMafg1-expressing cells. (d) M10M3 and (e) M167M1 cells harboring TRE-Cas9 and sgGFP or sgMafg1 were plated at a low density, and colony-forming ability was examined, which is shown as percent surface area covered by colonies. (f) M10M3 and (g) M167M1 cells harboring TRE-Cas9 and sgGFP or sgMafg1 were plated in soft agar, and anchorage-independent growth was examined. Dox, doxycycline; sgGFP, sgRNA targeting GFP; sgMafg1, sgRNA targeting Mafg.
Fig 2: EIF3J-AS1 is positively correlated with MAFG in PCa. A, GEPIA assessed EIF3J-AS1, and MAFG was positively correlated in PCa tissues. B and C, MAFG mRNA and protein expression in PC-3 and DU145 cells transfected with si-EI3J-AS1 was evaluated by qRT-PCR and western blot assay. D, qRT-PCR was used to assess EIF3J-AS1 expression in PCa cells (DU145, PC3 and LNCaP) and the human normal prostate epithelial cell line RWPE-1. E, MAFG mRNA expression in clinical tissue was assessed by qRT-PCR. F, The correlation between MAFG expression and EI3J-AS1 expression in 36 PCa tissues was evaluated by Pearson's correlation analysis. *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 3: Knockdown of MAFG inhibits PCa cell proliferation and invasion in vitro. A and B, qRT-PCR and WB assays assessed MAFG mRNA and protein expression in the indicated treated PCa cell lines. C, The proliferation of the indicated PCa cells was determined by CCK-8 assay. D, The migration of the indicated PCa cells was determined by Transwell assay without Matrigel-coated membranes. E, The invasion of the indicated PCa cells was determined by Transwell assay with Matrigel-coated membranes. *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 4: Pathway enrichment of TF downstream genes and matching to key signalling pathways of AS or vascular endothelial cell apoptosis. The transcription factors STAT6 (a), ATF3 (b), JUN (c), JUNB (d), MAFF (e), MAFG (f), and STAT1 (g) were enriched in known key signalling pathways related to atherosclerosis. MAFF (e) and MAFG (f) in red letters are novel candidates of TFs involved in vascular endothelial cell apoptosis and were enriched in known key signalling pathways of atherosclerosis. Signalling pathways with bold letters are known key signalling pathways of atherosclerosis. e, f The transcription factors indicated with red letters are novel candidate key TFs involved in vascular endothelial cell apoptosis
Fig 5: EIF3J-AS1 enhances PCa progression by upregulating MAFG A. qRT-PCR assays assessed MAFG mRNA expression in the indicated treated PC cell lines. B, Proliferation in the indicated PCa cells was determined by CCK-8 assay. C, The migration of the indicated PCa cells was determined by Transwell assay without Matrigel-coated membranes. D, The invasion of the indicated PCa cells was determined by Transwell assay with Matrigel-coated membranes. *P < 0.05, **P < 0.01, ***P < 0.001
Supplier Page from Abcam for Anti-MAFG antibody