Journal: International Journal of Molecular Sciences

Article Title: DNA Damage-Response Pathway Heterogeneity of Human Lung Cancer A549 and H1299 Cells Determines Sensitivity to 8-Chloro-Adenosine

doi: 10.3390/ijms19061587

Figure Lengend Snippet: Effects of 8-Cl-Ado on cell growth and apoptosis. A549 and H1299 cells were exposed to 2 μM 8-Cl-Ado for indicated hours. ( A ) Cell proliferation was evaluated with MTT assay (see materials and methods). Data represent mean ± SD ( n = 3); ( B ) cells exposed to 8-Cl-Ado for 48 h were stained with propidium iodide whose signal was measured by FACScan. Apoptotic cells (subG1/<2N) were assayed by the computer program CELLQuest. Data are representative of three independent experiments; ( C ) a representative Western blotting for Procaspase-3 activation and PARP-1 cleavage in 8-Cl-Ado-exposed cells. β-Actin as a loading control; ( D ) relative levels of Procaspase-3, Procaspase-3-cleaved fragments (p21 and p17), PARP-1 (p115) and its cleaved product (p85) in Western blotting. The blots were screened/quantified with the software Quantity One (Bio Rad) and normalized against β-Actin level, and the ratio of target protein to Actin from control (0 h exposure) cells was designated as “1” (100%). Data represent mean ± SD ( n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Apoptosis was assayed by the appearance of a sub-G1 (<2N ploidy) population by the computer program CELLQuest (Becton Dickinson, Franklin Lakes, NJ, USA).

Techniques: MTT Assay, Staining, Western Blot, Activation Assay, Control, Software