Figure 5
ABCA1 suppresses a mitochondria-dependent apoptosis pathway. (a–c) Neuro-2a cells were transiently transfected with either vehicle or Abca1, siNC, and siAbca1 expression constructs. For OGD, cells were detached from plates 24 h after transfection and reseeded. Twenty-four hours after reseeding, cells were subjected to 8 h of OGD and 1 h of reperfusion. mRNA expression of Abca1, Bax, and Bcl2 was measured by RT-qPCR (n = 3). Cell viability was determined by CCK-8 assay. (d) Caspase-3, -8, and -9 activation was assessed by Western-blot analysis. Representative results are illustrated, and values from densitometric analyses are shown after normalization to α-tubulin from the controls (n = 3). Full-length blots are presented in Supplementary Fig. 4 (e) Mitochondrial membrane potentials were determined by flow cytometry analysis using JC-1 staining. Representative results are illustrated, and values are reported mean ± standard deviation (n = 4). Statistical analyses were performed using t-tests (* = p < 0.05, ** = p < 0.01, *** = p < 0.001). Abbreviations: Abca1, ATP-binding cassette subfamily A member 1; siNC, non-targeting control small interfering ribonucleic acid; siAbca1, Abca1 siRNA; OGD, oxygen–glucose deprivation; mRNA, messenger ribonucleic acid; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; Bax, Bcl2 associated X; Bcl2, B-cell lymphoma 2; JC-1, 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-benzamidazolocarbocyanin iodide.
