Fig 2: Effect of CDK10 on cell growth and proliferation. (A) H522 cells were transfected with the indicated siRNAs, and detected by western blotting. (B) Cell growth assay was performed to measure the H522 cell growth rate in different groups. Mean ± SD, t test, ***P < 0.001. (C) H522 cells transfected with indicated siRNAs were cultured for two weeks, and the colonies were counted. Mean ± SD, t test, ***P < 0.001. (D) H522 cells transfected with indicated siRNAs were labelled with DAPI and EdU, and the percentage of EdU-positive cells was calculated. Scale bar, 50 mm. Mean ± SD, t test, ***P < 0.001. (E) A549 cells were transfected with the indicated siRNAs, and detected by western blotting. (F) EdU-positive cells were detected in A549 cells, transfected with the indicated siRNAs. Scale bar, 50 mm. Mean ± SD, t test, ***P < 0.001.

Fig 3: MAPK8, CDK10, and GSK3β are associated with exosomes: (A) Schematic diagram of the differential ultracentrifugation and size exclusion chromatography exosome isolation methods. (B) U1 exosomes and (C) ACH2 exosomes isolated via differential ultracentrifugation are loaded onto IZON 35 nm qEV original exclusion columns. Forty fractions are collected and pooled in sets of 5. Pooled fractions are then probed for MAPK8, CDK10, GSK3β, HSP90, and CD63 using Western blot. This figure is representative of a data set of n = 1.

Fig 4: Analysis of CDK10 expression and clinical prognosis in lung cancer. (A) The gene expression level of CDK10 in normal and lung adenocarcinoma tissues in the TCGA database. Mean ± SD, t test, *P < 0.05. (B) Western blotting analysis of CDK10 in HBE, LL2, and eight human lung cancer cell lines. (C) Immunohistochemical stain images of CDK10 in lung cancer and adjacent tissues. Scale bar, 50 mm. (D) Line and scatter plots of CDK10 protein expression scores in 12 cases of lung cancer and paired adjacent tissues. Mean ± SD, t test, *P < 0.05. (E) Kaplan-Meier survival curves show the relationship between CDK10 expression levels and clinical prognosis in lung cancer patients (https://kmplot.com/analysis/). The cutoff value used in analysis is 235, and the expression range of the probe is (5-4,896). HR: 0.82, Logrank P = 0.001.

Fig 5: CDK10 is present inside ACH2 exosomes: (A) Schematic illustration of proteinase protection assay adapted from [39], in which dark blue dots represent intact proteins and light blue dots represent degraded proteins. (B) ACH2 exosomes were treated with or without Triton X-100 in the presence of proteinase K. Exosomes were then immunoprecipitated with antibodies against CDK10 and used in a kinase assay. (C) Purified CDK5 kinase was used as a positive control. This figure represents data of n = 1.