Fig 2: Andrographis and berberine mutually enhance the inhibition of DNA replication in mouse xenografts and patient-derived tumor organoids. (A) A bar graph showing the relative mRNA expression of FEN1, MCM7, PRIM1, MCM5, POLA1, MCM4, and PCNA in dissected tumor xenografts after being treated with berberine and/or Andrographis; (B) A bar graph showing the relative mRNA expression of DNA-replication-related genes in two strains of patient-derived tumor organoids; (C,D) The protein immunoblot analysis of DNA-replication-related proteins in RKO cells and dissected tumor xenografts treated with berberine and/or Andrographis. Statistical Significance: * p < 0.05, ** p < 0.01, *** p < 0.001. BBR, berberine; Andro, Andrographis.

Fig 3: FEN1 serves as a direct miR-134-3p target in SKOV-3 and OVCAR-3 cells. (A) The putative binding site between FEN1 and miR-134-3p was predicted by starBase. (B) Dual-luciferase reporter assay was performed to reveal the association between miR-134-3p and FEN1. (C) Reverse transcription-quantitative PCR was performed to assess the mRNA expression of FEN1. (D) Western blot analysis was employed to assess the protein expression levels of FEN1 in SKOV-3 and OVCAR-3 cells transfected with miR-134-3p mimic. The data are presented as the mean ± SD (n=3). **P<0.01 vs. NC mimic. miR, microRNA; NC, negative control; FEN1, flap structure-specific endonuclease 1; WT, wild-type; MUT, mutant.

Fig 4: Overexpression of FEN1 reverses the effects of miR-134-3p on SKOV-3 and OVCAR-3 cells. SKOV-3 and OVCAR-3 cells were co-transfected with miR-134-3p mimic or NC mimic along with Lv-FEN1 or Lv-NC, respectively. (A) Reverse transcription-quantitative PCR was performed to assess FEN1 expression. (B) Cell Counting Kit-8 was performed to assess cell viability at 0, 24, 48 and 72 h. (C) Colony formation was performed to assess colony formation of the cells (magnification, ×40). (D) TUNEL (scale bar, 200 µm) and (E) Transwell assays (scale bar, 100 µm) were performed to assess the apoptosis and invasion of the cells, respectively. The data are presented as the mean ± SD (n=3). *P<0.05 and **P<0.01 vs. NC mimic+Lv-NC. #P<0.05 and ##P<0.01 vs. miR-134-3p mimic+Lv-NC. miR, microRNA; NC, negative control; FEN1, flap structure-specific endonuclease 1; OD, optical density; Lv, lentiviral expression vector (pcDNA3.1).