Journal: International journal of molecular sciences

Article Title: Glioma Stem Cells Are Sensitized to BCL-2 Family Inhibition by Compromising Histone Deacetylases.

doi: 10.3390/ijms241813688

Figure Lengend Snippet: Figure 4. Replacing MEKi with BCL-2 family inhibitors triggers apoptosis. (A) GSC12 was treated for 24 h with the indicated combinations: MEKi (25 nM trametinib), HDACi (2.5 µM vorinostat), NX (100 nM navitoclax) followed by analysis of apoptosis (CC3). (B) GSC12 was treated for 24 h with the indicated combinations: MEKi (25 nM trametinib), HDACi (2.5 µM vorinostat), NX (100 nM navitoclax), MCL1i (100 nM S63845) followed by analysis of apoptosis (CC3). Actin protein was used as a loading control for (A–C). GSC39 cells were treated for 24 h with the indicated combinations: MEKi (25 nM trametinib), HDACi (2.5 mM vorinostat), MCL1i (100 nM S63845) followed by analysis for apoptosis by annexin V-propidium iodide (PI) flow cytometry measuring apoptosis (annexin V positive/PI negative, Q3). Graph depicts mean +/−SEM, (n = 7). (D) Diagram summarizing results highlighting the use of HDACi to interfere with GBM cell cycle combined with potentiating intrinsic apoptosis using either MEKi to rescue BH3-only proteins, or inhibitors of pro-survival BCL-2 family proteins. Graphs portray means + SEM. p = * ≤0.05; *** ≤0.001, t-test.

Article Snippet: The cells were stained with 400 μL of staining solution composed of 0.1% v/v Triton-X 100 in PBS, 2 mg DNAse-free RNase A (Thermo Fisher EN0531), and 500 μg/mL propidium iodide (Biogems 60910-00), and incubated for 15 min at room temperature.

Techniques: Control, Cytometry