Extended Data Fig. 1: KSHV vBcl-2 targets NM23-H2 independently of inhibition of autophagy and apoptosis.
a, Co-immunoprecipitation (co-IP) of vBcl-2 with NM23-H2 in HEK293T cells co-transfected with HA-vBcl-2 and Flag-NM23-H2. Actin serves as a loading control. b, Co-IP of endogenous NM23-H2 with vBcl-2 in HEK293T cells expressing empty vector (Vec) or HA-vBcl-2. c, Schematic of WT vBcl-2 and deletion mutants, with binding to NM23-H2 determined by co-IP of Flag-NM23-H2 with HA-vBcl-2 in HEK293T cell lysates. BH (Bcl-2 homology) domains and the conserved 84W85G86R residues in BH1 domain are highlighted. TM, transmembrane domain. +, strong binding; -, no binding. d, Co-IP of Flag-tagged NM23-H2 with HA-tagged wild-type (WT) or mutant vBcl-2 in HEK293T cells. The binding results are summarized in c. e, IB analysis of LC3-I/LC3-II and p62 in HEK293T cells stably expressing Vec or HA-vBcl-2 (WT or mutants) with or without Torin 1 (50 nM, 3 h). f, Densitometric quantification of LC3-II/LC3-I (red bars) and p62/actin (blue) from e. n = 3. g, Flow cytometry analysis of apoptosis in HEK293T cells stably expressing Vec or HA-vBcl-2 (WT or mutants) treated with TNF-α (10 ng/ml) and CHX (5 μg/ml) for 4 h. Early apoptotic cells (Annexin V-positive and PI-negative) were quantified (n = 3). Data in a,b,d,e are from one experiment that is representative of three independent experiments. See Source Data for uncropped data of a,b,d,e. Data shown in f,g are mean ± s.d. analyzed by one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. *, p < 0.05; **, p < 0.01; ****, p < 0.0001; ns, not significant. Exact P values are provided in Source Data Extended Data Fig. 1.
