Fig 2: Cdk2 specifically inhibited the expression of IL‐23. RAW264.7 cells were infected with retrovirus carrying Cdk2 shRNA for 72 h. (A,B) qPCR and Western blot assay for the mRNA and protein levels of Cdk2, respectively. (C) Cdk2‐silencing RAW264.7 cells were treated with LPS (100 ng/ml) for 2 h, and inductions of multiple proinflammatory cytokines were measured by qPCR. Actin was used as loading control and for the relative normalization. (D) The chemical structure of CDKs inhibitors, AT7519. (E) BMDMs were generated from the WT B6 mice with M‐CSF (50 ng/ml) for 5 days. After pretreated with AT7519 (50 nM) for 24 h, these BMDMs were stimulated with LPS (100 ng/ml) for 2 h, and inductions of multiple proinflammatory cytokines were measured by qPCR. Data were shown as means ± SD based on three independent experiments at least. Two‐tailed Student's t‐tests were performed. BMDM, bone marrow‐derived macrophages; CDK, cyclin‐dependent kinase 2; DMSO, dimethyl sulfoxide; IL‐23, interleukin‐23; LPS, lipopolysaccharides; M‐CSF, macrophage colony‐stimulating factor; mRNA, messneger RNA; Nos2, nitric oxide synthase 2; qPCR, quantitative polymerase chain reaction; shRNA, short hairpin RNA; Tnf, tumor necrosis factor; WT, wild‐type. *p < .05 and **p < .01.

Fig 3: MORC2 silencing inhibits the proliferation of U251 cells. (A) Cell viability was assessed using a CCK-8 assay. (B and C) Ki67 expression was detected by immunofluorescence staining and quantification. Scale bar, 50 µm. (D) PCNA, CDK2 and Cyclin E1 expression was measured by western blotting. The grouping of images from different parts of the same gel. *P<0.05, **P<0.01 and ***P<0.001 vs. shRNA-NC. Sh, short hairpin; MORC2, Microrchidia family CW-type zinc finger 2; CDK2, cyclin-dependent kinase 2; NC, negative control; PCNA, proliferating cell nuclear antigen; control, untransfected U251 cells.

Fig 4: Effects of EI24 knockdown or overexpression on cell cycle progression in esophageal squamous cell carcinoma (ESCC) cells. Cell-cycle analysis revealed that ectopic expression of EI24 in EC9706 and EC109 cells increased the percentage of cells in the G1 phase and decreased the percentage of cells in S phase (A), while knocking down EI24 expression in TE-1 cells decreased the percentage of cells in the G1 phase and increased the percentage in S phase (B). (C) Western blot analysis of the expression of CDK2, CDK4, and cyclin D1 in ESCC cells. CDK2, CDK4, and cyclin D1 expression were decreased in EI24-OE cells compared with EI24-NC cells. EI24 depletion reversed this effect on CDK2, CDK4, and cyclin D1 expression. β-Actin was used as an internal control. *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 5: Overexpression of DSCR1-4 showed tumor suppressive function in CRC cells in vitro. a Western blot showed that overexpression of DSCR1-4 increased p21, p16, and hyperphosphorylation of NFAT1 and NFAT2, while decreased cyclin D1, CDK2, and CDK4. b CCK8 assays showed that overexpression of DSCR1-4 inhibited cell proliferation of CRC cells. Data are mean ± SEM of 5 replicates. c Overexpression of DSCR1-4 inhibited cell colony formation of CRC cells. Data are mean ± SEM of 3 replicates. d Overexpression of DSCR1-4 induced senescence of CRC cells by SA-β-gal staining. Two-tailed Student t test, *p < 0.05, **p < 0.01, ***p < 0.001