Fig. 4.

Truncated protein P5 interacted with BECN1. a, b N2a cells were cotransfected with Flag-P5 and Myc-BECN1 (a) or singly transfected with siBecn1 RNA for 12 h and infected with RABV strain HEP-Flury at MOI = 2 for 24 h (b), and the viral P/P5 protein (green), BECN1 (red), and DAPI (blue) were detected using the indicated antibodies via confocal microscopy. White arrows indicate the colocalization of the ring-like structure. Scale bar: 10 μm. The graph shows the quantification of the percentage of BECN1 localization with P5 ring-like structure. c HEK 293 T cells were cotransfected with the plasmids containing the truncated P genes and Myc-BECN1 for 48 h, and the interactions between the truncated P protein and BECN1 were determined using the indicated antibodies. IP, immunoprecipitation. The asterisk indicates the heavy chains. d Schematic diagram of the full-length and truncated BECN1 proteins. e N2a cells were singly transfected or cotransfected with the plasmids encoding the truncated BECN1 genes and Flag-P for 24 h, and the viral P protein (red), BECN1 (green), and DAPI (blue) were detected using the indicated antibodies via confocal microscopy. White arrows indicate the colocalization of the ring-like structure. Scale bar: 10 μm. The graph shows the quantification of the ring-like structures formed with BECN1 deletion mutants. f HEK 293 T cells were cotransfected with the plasmids containing the truncated Becn1 genes and Flag-P for 48 h, and the interactions between the truncated BECN1 proteins and P were determined using the indicated antibodies. IP, immunoprecipitation. The asterisk indicates the heavy chains. Means and SD (error bars) of three independent experiments are indicated (***, P < 0.001)