Journal: Heliyon

Article Title: Lnc-PLA2G4A-4 facilitates the progression of hepatocellular carcinoma by inducing versican expression via sponging miR-23b-3p

doi: 10.1016/j.heliyon.2023.e18698

Figure Lengend Snippet: Lnc-PLA2G4A-4 directly binds to miR-23b-3p. A. Fluorescence in Situ Hybridization assay used to examine the expression and location of Lnc-PLA2G4A-4 in SMMC-7721 and SK-Hep-1 cells. B. Cell fractionation assay used to examine the location of Lnc-PLA2G4A-4 in SMMC-7721 and SK-Hep-1 cells. C. The predicted results of Lnc-PLA2G4A-4 downstream target genes from three databases. D. Relative luciferase activities in 293T cells co-transfected with the mimic-NC/miR-567 mimic/miR-888 mimic/miR-23b-3p mimic and Lnc-PLA2G4A-4-WT reporter vector. E. Up: prediction of miR-23b-3p binding sites in Lnc-PLA2G4A-4 (Lnc-PLA2G4A-4-WT) and the design of Lnc-PLA2G4A-4 mutation sequence (Lnc-PLA2G4A-4-Mut). Down: luciferase reporter assays were performed in 293T cells co-transfected with the miR-23b-3p mimic and Lnc-PLA2G4A-4-WT or Lnc-PLA2G4A-4-Mut reporter vector. F. Relative expression of miR-23b in different HCC cell lines. G. Expression of miR-23b-3p in 50 normal tissue and 374 HCC tumor tissue from TCGA cohort in Student's unpaired t -test. H. Expression of miR-23b in 49 normal tissue and 49 HCC tumor tissue from TCGA cohort in Student's paired t -test. I. Relative expression of miR-23b in Lnc-PLA2G4A-4 over-expression cells or Lnc-PLA2G4A-4 knockdown cells, respectively. J. Correlation between miR-23b-3p and Lnc-PLA2G4A-4 expression in HCC tissues of MCHH. K. Correlation between miR-23b-3p and Lnc-PLA2G4A-4 expression in HCC tissues of TCGA.

Article Snippet: Human HCC cell line SNU398, SNU449, SK-Hep-1, Hep3B, HepG2/C3A, as well as Human Embryonic Kidney 293T (HEK-293T) cell line were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Fluorescence, In Situ Hybridization, Expressing, Cell Fractionation, Luciferase, Transfection, Plasmid Preparation, Binding Assay, Mutagenesis, Sequencing, Over Expression

Journal: Cancer Science

Article Title: Targeting phosphomevalonate kinase enhances radiosensitivity via ubiquitination of the replication protein A1 in lung cancer cells

doi: 10.1111/cas.15896

Figure Lengend Snippet: PMVK depletion decreases RPA1 protein level and inhibits HR repair. (A) Cells were transfected with the indicated siRNAs for 24 h. The cell lysates were subjected to western blotting. Equal loading of the protein samples was confirmed by western blotting of α‐tubulin. (B) Cells were transiently transfected with Flag‐RPA1 for 24 h, and then treated with siPMVK and RT. Overexpression of RPA1 and suppression of PMVK expression was confirmed by western blotting. Equal loading of the protein samples was confirmed by western blotting of α‐tubulin. The cells were incubated for 9 days to allow colony formation. Surviving colonies were counted after staining with crystal violet. Columns show the mean of three experiments; SE. * p < 0.001. (C) Cells were co‐transfected with Flag‐RPA1 and HA‐ubiquitin plasmids for 24 h, and then transfected with indicated siRNAs for another 24 h. MG132 was added for 4 h before harvesting, and cells were collected for immunoprecipitation (IP) with Flag‐agarose beads and then subjected to western blotting analysis. Equal loading of the protein samples was confirmed by western blotting of α‐tubulin. (D) HR and NHEJ repair efficiency determined with the DR‐GFP and EJ5‐GFP reporters in 293T cells transfected with the indicated siRNAs using flow cytometric analyses. siRAD51 and siDNA‐PK were positive controls of HR and NHEJ, respectively (upper). Protein expression of Rad51, DNA‐PK, PMVK, and RPA1, was detected by western blotting. Equal loading of the protein samples was confirmed by western blotting of α‐tubulin (bottom). Columns show the mean of three experiments; SE. * p < 0.001, compared with the siCon‐transfected cells.

Article Snippet: Human lung cancer (A549, H1299, H460, and EKVX), normal mouse lung (MLE‐12), normal human lung (MRC5), and human embryonic kidney (293T) cell lines were obtained from the ATCC.

Techniques: Transfection, Western Blot, Over Expression, Expressing, Incubation, Staining, Immunoprecipitation