Fig 2: Low dose androgen biology is not driven by the canonical transcriptional activity ascribed to AR.Heatmap of ATAC-seq signal within a 6 kb window of all high-confidence differentially accessible peaks identified in response to indicated androgen dose compared to vehicle treated VCaP (a) or LNCaP (b) cells. Differentially accessible peaks were subdivided based on whether they are significantly increased (VCaP Up or LNCaP Up) or significantly decreased (VCaP down or LNCaP down). Quantification of unique and common high-confidence differentially accessible peaks identified in response to indicated androgen dose compared to vehicle treated VCaP (c) or LNCaP (d) cells. e Representative genomic tracks of ATAC-seq sites within classical AR regulated genes (i.e., KLK3, FKBP5 and STEAP4) or LD regulated genes (i.e., E2F1, CDC6 and FOXM1) in response to vehicle (0) or increasing androgens in VCaP cells.

Fig 3: FoxM1 is a downstream target of miR-134, which directly binds to the 3′UTR of FoxM1. (A) Putative miR-134-binding sites in the 3′UTR of FoxM1 mRNA. A mutation was generated in the FoxM1 3′UTR sequence in the complementary site for the seed region of miR-134. (B) Relative expression levels of FoxM1 in GC tissues without H. pylori infection, and tissues infected with H. pyloriCagA−, H. pyloriCagA+/P− and H. pyloriCagA+/P+. (C) A negative correlation was observed between FoxM1 mRNA and miR-134 expression in GC tissues infected with H. pyloriCagA+/P+. (D) Expression levels of miR-134 in SGC-7901 cells transfected with inhibitor control or miR-134 inhibitors, as determined by RT-qPCR analysis. (E) mRNA expression levels of FoxM1 were analyzed by RT-qPCR analysis in SGC-7901 cells transfected with mimic control, miR-134 mimics, inhibitor control or miR-134 inhibitors. GAPDH was used as an internal control. (F) Protein expression levels of FoxM1 were determined by western blotting in the groups. GAPDH was used as an internal control. (G) Luciferase assay in 293T cells co-transfected with miR-134 mimics or mimic control and a luciferase reporter containing wild type or mutant FoxM1 3′UTR. (H) Immunofluorescence analysis revealed the alteration of FoxM1 in H. pyloriCagA+/P+-infected SGC-7901 cells transfected with miR-134 mimics or mimic control. Images merged with DAPI are presented. Scale bar, 50 µm. *P<0.05 vs. the mimic control group; #P<0.05 vs. the inhibitor control group. 3′UTR, 3′-untranslated region; CagA, cytotoxin-associated gene A; H. pyloriCagA-, CagA-negative H. pylori; H. pyloriCagA+/P−, CagA-positive and PBP1A mutation-negative H. pylori; H. pyloriCagA+/P+, CagA- and PBP1A mutation-positive H. pylori; FoxM1, Forkhead box M1; H. pylori, Helicobacter pylori; miR-134, microRNA-134; PBP1A, penicillin-binding protein 1A; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Fig 4: Circ-0039411 affected proliferation, migration, and EMT relying on FOXM1 in LUAD.a The qRT-PCR and western blot assays was utilized to detect the effect of circ-0039411 inhibition on FOXM1 expression. GAPDH was the internal control. b, c Cell proliferation was detected by EdU (scale bar = 100 μm) and colony formation assays. d Transwell assay was utilized to test the cell migration (scale bar = 100 μm). e Western blot assay was adopted to detect EMT-related proteins. GAPDH was the internal control. **P < 0.01. n.s. meant no significance.

Fig 5: Circ-0039411 transcription was activated by FOXM1 in LUAD.a The result of qRT-PCR of circ-0039411 expression in LUAD and normal cells. b qRT-PCR was utilized to test RNA expression in LUAD cells after Act D treatment. c The qRT-PCR detection of RNA expression after LUAD cells were treated with RNase R. d Ring forming diagram of circ-0039411. e AGE (agarose gel electrophoresis) results of PCR products amplified by convergent or divergent primers. f The binding motif of FOXM1 and the two possible FOXM1 binding sites in circ-0039411 promoter. g Luciferase reporter assay was carried out to estimate the impact of FOXM1 on circ-0039411 transcription. h qRT-PCR measured the enrichment of circ-0039411 promoter in ChIP products of FOXM1. *P < 0.05, **P < 0.01.