Fig 2: Regulation of the c-Myc-CDK4/6-phosphoRb axis by UHRF1 in B-ALL and T-ALL. A knockdown of UHRF1 led to reduced c-Myc protein in (A) T-ALL and (B) B-ALL cells. The ALL cells were transfected with siRNA-targeting UHRF1 and a control siRNA. After being incubated for 48 h, the UHRF1, c-Myc, CDK4/6, and p-RB/RB protein levels were analyzed using Western blot and densitometry. The uncropped blots are shown in Figure S1.

Fig 3: Inhibition of YAP induces cell cycle arrest in RPMI 2650 cells. (A) RPMI 2650 cells treated with VP (0, 10, 15, 20 μM) after 24 h and subjected to cell cycle detection. (B) The proportion of cells in each phase was shown (n = 3 for each group, one-way ANOVA). (C) Representative Western blotting analysis of cell cycle progression proteins cyclinD1 and CDK4 in RPMI 2650 cells treated with VP for 24 h (n= 3 for each group). (D) RPMI 2650 cells were treated with VP (0, 10, 15, 20 μM) for 24 h and the apoptosis was detected (n= 3 for each group). (E) The proportion of apoptotic cells treated with VP of different concentrations after 24 h was shown (n = 3 for each group, one-way ANOVA). Data were presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 4: TRAF3 is a direct target of miR‐361‐3p. (A) Venn diagram of predicted miR‐361‐3p targets by five programs (miRDB, DIANA, miRwalk, TargetScan and miRTarBase). (B) The seed regions of miR‐361‐3p, the seed‐recognising sites in the TRAF3 3′ UTR, and the nucleotides mutated in TRAF3 mutant 3′ UTR are shown. (C) Luciferase reporter assay was conducted to verify that miR‐361‐3p bound to the 3′‐UTR region of TRAF3 directly. miR‐361‐3p overexpression significantly suppressed, while miR‐361‐3p loss increased the luciferase activity that carried wild‐type (WT) but not mutant (MUT) 3′‐UTR of TRAF3. (D) RIP assays confirmed the binding status between miR‐361‐3p and TRAF3 in CRC cell lines, respectively. (E) miR‐361‐3p overexpression decreased the level of TRAF3 mRNA and protein in CRC cells. (F) qRT‐PCR analysis indicated that the mRNA expression of TRAF3 in low‐miR‐361‐3p tumours was significantly higher than that in high‐miR‐361‐3p tumours. (G) Pearson's correlation analysis showed the negative correlation of TRAF3 with miR‐361‐3p. (H) Protein levels of HIF‐1α, TRAF3, ki‐67, CDK4, Cyclin D1, Bcl‐2 and Bax in CRC tissues were determined by IHC. Scale bar = 200 μm. FISH was used to detect the miR‐361‐3p in CRC tissues. Scale bar = 50 μm. (I) The relative expression of exosomal miR‐361‐3p in plasma in different tumour size. (J) Comparison of overall survival (OS) and disease‐free survival (DFS) between patients with high expression of exosomal miR‐361‐3p in plasma and low expressing cases in CRC via Kaplan–Meier analysis. Data represent the mean ± SD. Student's t‐test was used to determine statistical significance: *p < 0.05 and ***p < 0.001

Fig 5: Overexpression of miR-486 suppresses the expression of CDK4 and BCAS2. (A) Predicted consequential pairing of miR-486 and CDK4/BCAS2. (B) Dual-Luciferase assay (**p<0.01). (C) Expression of CDK4/BCAS2 was suppressed in the miR-486 mimic-transfected EC9706 cells (***p<0.001, **p<0.01).