Fig 1: CDKs inhibitor suppressed IL‐23 expression via regulating DCAF2 expression. (A) qPCR analysis of Dcaf2 in the pancreas from nontreated and pancreatitis mice. (B,C) 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 expression of Dcaf2 was measured by qPCR and Western blot. Actin was used as loading control and for the relative normalization. (D) BMDMs were pretreated with MLN4924 (5 nM) and AT7519 (50 nM) for 24 h. The expressions of Il23a and Tnf were evaluated by qPCR. (E) RAW264.7 cells were infected with retrovirus carrying DCAF2 expressing sequence and incubated with AT7519 (50 nM) for 24 h. The inductions of Il23a and Tnf were evaluated by qPCR. Data were shown as means ± SD based on three independent experiments at least. Two‐tailed Student t‐tests were performed. BMDM, bone marrow‐derived macrophages; CDK, cyclin‐dependent kinase 2; DCAF2, DDB1–cullin‐4‐associated factor‐2; DMSO, dimethyl sulfoxide; IL‐23, interleukin‐23; LPS, lipopolysaccharides; M‐CSF, macrophage colony‐stimulating factor; qPCR, quantitative polymerase chain reaction; Tnf, tumor necrosis factor; WT, wild‐type. *p < .05 and **p < .01.
Fig 2: The expression levels of DTL in different tissues and cancers. A The expression of DTL in 31 different tissues. B Comparisons of the expression of DTL levels between different cancer tissues and normal tissues, *P < 0.05, **P < 0.01, ***P < 0.001. C Uncovering the mutation frequency of DTL in 33 different types of tumors
Fig 3: The representative IHC and IF images of DTL and CD3 staining and the statistical map of them, Kaplan–Meier analysis,*P < 0.05, **P < 0.01, ***P < 0.001. A IHC images of DTL staining were showed that DTL was successively overexpressed in normal, low-grade, middle-grade and high-grade LIHC tissues, 20X. B IF images of DTL staining were displayed that DTL was overexpressed in LIHC, especially in cancer nests (DTL: red, DAPI: blue, 40X). C-DRepresentative IF images of DTL staining were displayed that DTL was overexpressed in BLCA and STAD (DTL: red, DAPI: blue, 20X). E Comparisons of the expression of DTL levels between cancer tissues and normal tissues in all liver, bladder and stomach clinical samples, (normal: blue, cancer: red). F Comparisons of the expression of DTL levels in different pathological stage in LIHC. G Comparisons of the expression of DTL levels in different original tumor stage (T) in BLCA. H Comparisons of the expression of Ki-67 levels in DTL high and low group of LIHC, (the group of DTL low expression: blue, the group of DTL low expression: red). I Representative images of the high and low CD3 + T cells infiltration groups in LIHC, BLCA and STAD were showed (CD3: green, DAPI: blue, 20X). J Comparisons of the percentage of CD3.+ T cells infiltration in DTL high and low group were carried out in LIHC, BLCA and STAD (normal: blue, cancer: red)
Fig 4: DTL was a key gene and an excellent diagnostic biomarker in BCa. (a) The scatter plot of all genes in the brown module; those circles located in the upper right indicate the key genes in these modules. (b) The protein-protein interaction network identified top 10 hub genes based on the STRING database. (c) ROC curve analysis of the selected top 10 hub genes in GSE13507. (d) DTL diagnosis ROC curve analysis in GSE13507.
Fig 5: Relationship between somatic mutations and DTL expression in HCC. (A) Somatic mutations in DTL-high and DTL-low expression groups. (B) Comparison of mutations between the high expression group and low expression group of DTL. *P < 0.05, **P < 0.01, ***P < 0.001.
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