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oai:figshare.com:article/34049147 2026-10-01T17:43:04Z category_4 category_12 category_13 category_14 category_64 category_128 category_132 category_134 portal_5 item_type_1 month_year_10_2026

NOXA KO restores cellular viability and decreases apoptosis during MARCH5 KO. Prasanth Viswanathan (10314782) Justine R. Bersonda (25157885) Jackson Gill (25157888) Alyssandra Navarro (25157891) Allison C. Farrar (25157894) Thomas Caldarera (25157897) Andrew Murphy (341397) Daniel Dunham (25157900) Karl W. Boehme (11384016) Mark Manzano (25157903) Biochemistry Cell Biology Genetics Molecular Biology Cancer Hematology Infectious Diseases Virology systems biology approach regulating mitochondrial homeostasis primary effusion lymphoma immunoprecipitation experiments show endogenous reciprocal co emerging clinical relevance cell viability due newly appreciated interaction causes several malignancies unexpected genetic interaction findings thus provide div >< p marchf5 oncogenes may pel cells suggesting mcl1 motivated us genetic interaction tumor cells causes resistance thus investigated therapeutic potential survival role significant increase protein complex previously shown pel ). mechanistic link kaposi ’ fitness advantage e3 ligase drug development dimerization activities crispr knockout combination therapies chemotherapy regimens attractive target associated herpesvirus apoptotic signal apoptotic activities antiviral signaling A. Workflow of the characterization experiments of the NOXA-MARCHF5 double KO (DKO) BC-1 Cas9 cells. Cells were transduced with a lentivirus encoding the targeting gRNA (SAFE4501 or NOXA g1), an eGFP reporter, and puromycin resistance gene (puroR). After selection and confirmation of NOXA KO by Western blotting (panel B), these cells were transduced with a second lentivirus encoding another targeting gRNA (gAAVS1, MARCHF5 g2 or g3) and the hygromycin resistance gene (hygR). After 7 days of selection (200 µg/mL) and confirmation of NOXA 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 NOXA g1 cells. C. Western blot of NOXA in MARCHF5-NOXA DKO cells. D-F. DKO cells were characterized using D. CellTiter-Glo2.0 (viability), E. Caspase-Glo 3/7 Assay (caspase 3/7 activity), or F. FCA (fitness). Caspase-Glo3/7 luminescence signals were normalized to CellTiter-Glo2.0 signals to normalize for total viable cells. All data in panels D-F, were further normalized to the SAFE4501-gAAVS1 DKO cells to establish the baseline. Statistical differences were calculated using one-way ANOVA with Dunnett’s multiple comparison post-hoc test (n = 3). Error bars, standard error of the mean. 2026-10-01T17:54:28Z Image Figure 10.1371/journal.ppat.1014665.g009 https://figshare.com/articles/figure/_p_i_NOXA_i_KO_restores_cellular_viability_and_decreases_apoptosis_during_i_MARCH5_i_KO_p_/34049147 CC BY 4.0