Journal: Genes

Article Title: Enhancing Transcriptional Reprogramming of Mesenchymal Glioblastoma with Grainyhead-like 2 and HDAC Inhibitors Leads to Apoptosis and Cell-Cycle Dysregulation

doi: 10.3390/genes14091787

Figure Lengend Snippet: Exogenous GRHL2 expression alters the GBM cell cycle. ( A ) Inducible LN229 cells were treated with dox for 72 h with or without HDACi for the last 24 h and were immunoblotted for indicators of cell-cycle phases pCDC2, pRB, cyclin B1, and p21. GAPDH was used as a loading control for protein expression. ( B ) Quantification of data in A. ( C ) Cell-cycle (propidium iodide) flow cytometry of parental or inducible LN229 treated with dox for 72 h. Graph displays mean values for G1, S, and G2/M phase. ( D ) Cell-cycle (propidium iodide) flow cytometry of iLN229 treated with dox for 3 or 14 days. Arrow indicates > 4 N cells. ( E ) Control or dox-treated (21 days) iLN229 cells were stained with CD44 to visualize the cell membrane and DAPI to visualize nuclei. Cells were then counted for multinucleated cells. White arrowheads identify multinucleated cells. Mean values of multinucleated cells are per 300 cells counted. Graphs depict means +/− SEM for n = 3. * p < 0.05 t -test; ** p < 0.01 t -test; *** p < 0.001 t -test.

Article Snippet: The cell pellet was resuspended in Annexin V Binding Buffer (BD Pharmingen, 556454 Franklin USA), and 5 × 10 5 cells were transferred to a FACS tube containing 5 µL propidium iodide (PI) staining solution (Peprotech 60910-00) and 5 µL FITC-Annexin V (BD Pharmingen, 556419).

Techniques: Expressing, Flow Cytometry, Staining, Membrane

Journal: Genes

Article Title: Enhancing Transcriptional Reprogramming of Mesenchymal Glioblastoma with Grainyhead-like 2 and HDAC Inhibitors Leads to Apoptosis and Cell-Cycle Dysregulation

doi: 10.3390/genes14091787

Figure Lengend Snippet: GRHL2 expression dysregulates cell cycle in glioma stem cells and associates with aneuploidy in patient tumors. ( A ) Inducible glioma stem cells (GSCs) were treated with dox for 72 h with or without HDACi for the last 24 and were immunoblotted for indicators of cell-cycle phases cyclin B1, and p21. β-Actin was used as a loading control for protein expression. ( B ) Cell-cycle (propidium iodide) flow cytometry of inducible GSCs treated with dox for 72 h. * p < 0.05, t -test. Arrow indicates > 4 N cells. ( C ) GRHL2 mRNA expression in patient tumors by cancer type, arranged by mean expression level and color-coded for aneuploidy score. ( D ) GRHL2 mRNA expression in cancers vs. aneuploidy score. ( n = 3077 scores < 6, n = 6579 > 6.) Data in C and D were captured from The Cancer Genome Atlas (TCGA) cBioBortal. **** p < 0.0001 t -test.

Article Snippet: The cell pellet was resuspended in Annexin V Binding Buffer (BD Pharmingen, 556454 Franklin USA), and 5 × 10 5 cells were transferred to a FACS tube containing 5 µL propidium iodide (PI) staining solution (Peprotech 60910-00) and 5 µL FITC-Annexin V (BD Pharmingen, 556419).

Techniques: Expressing, Flow Cytometry