Fig. 4: Dissection of the function of NPC1 in baculovirus endosomal escape.

DiOC18-staining experiment for illustration of NPC1 function in baculovirus membrane fusion, as analyzed by confocal microscopy (a) or flow cytometry analyses (b). The p-value between DiOC18 only and Nontarget groups is 1e-5. The p-value between NPC1^−/− and NPC1 rescued groups is 4e-6. c The structural organization of NPC1. d Design of NPC1 constructs. Analyses of the interactions between HA-tagged GP64 and Myc-tagged full-length NPC1, NPC1-A, NPC1-C, NPC1-I (e), NPC1-ΔA, NPC1-ΔC or NPC1-ΔI (f). g Investigation of the effects of overexpression rescue with full-length or truncation constructs of NPC1 on baculovirus transduction, as determined by the EGFP-positive cells via flow cytometry. Mock, PBS treatment without baculovirus. The p-values between NPC1^−/− and Nontarget and NPC1 rescued groups are 8e-8 and 1e-7, respectively. h Neutralization of Bac-EGFP transduction in HEK-293T cells by recombinant protein of NPC1 C ___domain, as determined by flow cytometry analyses. i ESI-MS confirmation of purified GST-I protein (n = 1). j Far-western analysis of the interaction between GP64 and GST-I. For (b, g, and h), Data are presented as mean ± SD (n = 3) from three biological replicates. The significant difference is analyzed by two-tailed unpaired Student’s t-test. For (e, f, and j), The experiment is repeated three times independently with similar results. Source data are provided as a Source Data file.