Journal: Neoplasia (New York, N.Y.)

Article Title: High p16 INK4A expression in glioblastoma is associated with senescence phenotype and better prognosis

doi: 10.1016/j.neo.2024.101116

Figure Lengend Snippet: In vitro Senescent GBM Model via p16 INK4A Overexpression Exhibit Senescence Phenotype. A The protein (upper panel) and mRNA expression levels (lower panel) of p16 INK4A ( CDKN2A ) in SNU201 after transduction with either an empty vector (Empty) or a p16 INK4A -overexpression vector (p16 INK4A ) is shown. kDa: kilodalton. NC: non-transduced SNU201. B The morphology of SNU201 cells post-transduction is depicted in the left panel, with SA-β-Gal staining results in the middle panel and quantification data on the right. C-D Real-time PCR data for representative SASPs and chemokines in SNU201 cells transduced with either vector. E-F IHC analysis for CCL2 and CCL13 in GBM tissues, with quantification data in the lower right panel. Statistical differences in ( A-D ) were analyzed using Student's t-test while those in ( E-F ) were analyzed using the Chi-square test.

Article Snippet: IHC was conducted using the following primary antibodies: p16 INK4A prediluted solution (805–4713; Roche, Tucson, AZ); αSMA, 1:200 (ab5694, Abcam, Cambridge, UK); CD68, 1:2500 (NBP2-48923, Novus Biologicals, Centennial, CO); GFAP, pre-diluted solution (IR524, Dako, Denmark); CD45, 1:1000 (ab8216, Abcam); CD11b, 1:4000 (ab133357, Abcam); CD3, 1:150 (ab135372, Abcam); Cleaved caspase-3, 1:400 (9661S, Cell Signaling Technology, Danvers, MA); CCL2, 1:100 (MAB2791, R&D Systems, Minneapolis, MN); CCL13 1:50 (sc-271124, Santa Cruz Biotechnology, Dallas, TX).

Techniques: In Vitro, Over Expression, Expressing, Transduction, Plasmid Preparation, Staining, Real-time Polymerase Chain Reaction

Journal: Neoplasia (New York, N.Y.)

Article Title: High p16 INK4A expression in glioblastoma is associated with senescence phenotype and better prognosis

doi: 10.1016/j.neo.2024.101116

Figure Lengend Snippet: The SASPs from Senescent GBM Cells Recruit Immune Cells in vitro . A The left panel shows a migration assay schematic, with quantification of migrated cells in the right panel (Empty: CM from empty vector-transduced SNU201; p16 INK4A : CM from p16 INK4A -overexpressed SNU201). B The left panel depicts a migration assay with si CCL2 or si CCL13 treatment in p16 INK4A -overexpressing SNU201, while the right panel shows CDKN2A (p16 INK4A ) mRNA expression levels. I The mRNA expression levels of CCL2 and CCL13 are shown in the left and right panels, respectively. J The left panel shows the number of migrated THP-1 cells with siControl or si CCL2 in p16 INK4A -overexpressed SNU201, and the right panel shows migrated Jurkat cells with siControl or si CCL13 . Statistical differences in ( A and C-D ) were analyzed using Student's t-test. NC: non-treated control; Empty: empty vector-transduced cells; p16 INK4A : p16 INK4A -overexpression vector-transduced cells. All graphs present mean ± standard deviation.

Article Snippet: IHC was conducted using the following primary antibodies: p16 INK4A prediluted solution (805–4713; Roche, Tucson, AZ); αSMA, 1:200 (ab5694, Abcam, Cambridge, UK); CD68, 1:2500 (NBP2-48923, Novus Biologicals, Centennial, CO); GFAP, pre-diluted solution (IR524, Dako, Denmark); CD45, 1:1000 (ab8216, Abcam); CD11b, 1:4000 (ab133357, Abcam); CD3, 1:150 (ab135372, Abcam); Cleaved caspase-3, 1:400 (9661S, Cell Signaling Technology, Danvers, MA); CCL2, 1:100 (MAB2791, R&D Systems, Minneapolis, MN); CCL13 1:50 (sc-271124, Santa Cruz Biotechnology, Dallas, TX).

Techniques: In Vitro, Migration, Plasmid Preparation, Expressing, Control, Over Expression, Standard Deviation

Journal: Neoplasia (New York, N.Y.)

Article Title: High p16 INK4A expression in glioblastoma is associated with senescence phenotype and better prognosis

doi: 10.1016/j.neo.2024.101116

Figure Lengend Snippet: The CCL13 expression is related to prognosis of GBM in TCGA dataset. A The survival graphs of GBM based on CCL13 expression (left panel) and CCL2 expression (right panel) are derived from the TCGA GBM-PanCancer dataset. B The schematic image illustrates how the presence of p16 INK4A -high GBM is linked to the tumor immune microenvironment and impacts patient prognosis.

Article Snippet: IHC was conducted using the following primary antibodies: p16 INK4A prediluted solution (805–4713; Roche, Tucson, AZ); αSMA, 1:200 (ab5694, Abcam, Cambridge, UK); CD68, 1:2500 (NBP2-48923, Novus Biologicals, Centennial, CO); GFAP, pre-diluted solution (IR524, Dako, Denmark); CD45, 1:1000 (ab8216, Abcam); CD11b, 1:4000 (ab133357, Abcam); CD3, 1:150 (ab135372, Abcam); Cleaved caspase-3, 1:400 (9661S, Cell Signaling Technology, Danvers, MA); CCL2, 1:100 (MAB2791, R&D Systems, Minneapolis, MN); CCL13 1:50 (sc-271124, Santa Cruz Biotechnology, Dallas, TX).

Techniques: Expressing, Derivative Assay