Fig 2: miR-144 is downregulated in PTC tissues and inversely correlates with E2F8 expression. a TCGA dataset showed that miR-144 was significantly downregulated in PTC tissues compared with adjacent normal tissues. b qRT-PCR analysis showed that miR-144 was significantly decreased along with more advanced T stage in PTC tissues. U6 was used as an internal control to normalize expression data. c Pearson correlation scatter plot of fold change in miR-144 levels and E2F8 mRNA levels in PTC tissues (n = 64). d A hypothetical model of the miR-144/E2F8/CCND1 axis controlling PTC cell proliferation. ***p < 0.001, # No significance

Fig 3: Influence of MYF5 on myogenesis via regulation of CCND1 expression. (A,B) C2C12 cells were transfected with MYF5 siRNA or Ctrl siRNA, along with a control vector [pcDNA3-Flag (pV)] or a plasmid vector that expressed Myc-tagged CCND1. Forty-eight hours later, the levels of CCND1 and MYF5 were analyzed by western blot analysis (A) and the degree of differentiation was analyzed by measuring creatine kinase activity at day 6 into differentiation (B). Data presented are the means and S.E.M. from four independent experiments; significance (P) is indicated. (C) Proposed model whereby MYF5 modulates myogenesis by acting upon CCND1 expression on two levels: first, MYF5 activates Ccnd1 transcription moderately, and second, MYF5 binds the Ccnd1 mRNA at the CR and 3'UTR, promoting Ccnd1 mRNA translation. The net effect is a CCND1-mediated increase in myoblast proliferation necessary at the initiation of myogenesis. N.S., not significant; *, P < 0.05; **, P < 0.01 (Student's t-test).

Fig 4: Effect of chondroitin polymerizing factor (CHPF) on global gene expression. A, Heatmap representation of 635 differentially expressed genes (DEGs) in A549 cells infected with shCHPF or shCtrl. P<0.05 and fold-change >2 (red: upregulated genes; green: downregulated genes). B, Functional pathway enrichment of the DEGs was analyzed via Ingenuity Pathway Analysis. The x-axis represents the pathway. Statistical significance is shown on the y-axis and represents the inverse log of the P-value. C, The network was built on the basis of microarray data of lung cancer cells. Red represents upregulation and green represents downregulation. D, Western blotting analysis was applied to evaluate the expression of CDH1, FOXM1, RRM2, HIF1A, CCND1, MKI67, and TNFRSF10B in shCHPF and shCtrl lung cancer cells. Data are reported as means±SD. **P<0.01 (Student's t-test).

Fig 5: MiR-18a affected primary myoblasts and RD cells proliferation.(A) Flow cytometry was used to analyze the percentage of cells in the G0/G1, S, and G2/M phases. n = 3, biological samples. **, p < 0.01. (B) The expression analysis for CCND1 and CDK6 in primary myoblasts transfected with miR-18a mimics or NC. GAPDH was used as an internal normalized reference. Data were shown as the means ± SEM (n = 4, biological samples). **, p < 0.01. (C) Western blot data showed the protein level of CCND1 and CDK6 in primary myoblasts transfected with miR-18a mimics or NC. Tubulin served as the internal normalized reference. (D) Flow cytometry was used to analyze the percentage of cells in the G0/G1, S, and G2/M phases. n = 3, biological samples. *, p < 0.05. (E) The expression analysis for CCND1 and CDK6 in RD cells transfected with miR-18a mimics or NC. GAPDH was used as an internal normalized reference. Data were shown as the means ± SEM (n = 4, biological samples). **, p < 0.01. (F) Western blot data showed the protein level of CCND1 and CDK6 in RD cells transfected with miR-18a mimics or NC. Tubulin served as the internal normalized reference.