<p><i>NOXA</i> KO restores cellular viability and decreases apoptosis during <i>MARCH5</i> KO.</p>

Cells were transduced with a lentivirus encoding the targeting gRNA (SAFE4501 or <i>NOXA</i> g1), an eGFP reporter, and puromycin resistance gene (puro^R). After selection and confirmation of <i>NOXA</i> KO by Western blotting (panel B), these cells were transduced with a second lentivirus encoding another targeting gRNA (g<i>AAVS1</i>, <i>MARCHF5</i> g2 or g3) and the hygromycin resistance gene (hyg^R).

After 7 days of selection (200 µg/mL) and confirmation of <i>NOXA</i> KO by Western blotting (panel C), cell densities were adjusted to 4 x 10⁵ cells/mL and seeded for CellTiter-Glo2.0 Assay, Caspase-Glo 3/7 Assay and FCA for 2 days. B. Western blot of NOXA in SAFE4501 or <i>NOXA</i> g1 cells. C. Western blot of NOXA in <i>MARCHF5</i>-<i>NOXA</i> DKO cells.