RABV infects sympathetic neurons in tri-chambers via axonal infection.

(A) Retrograde infection in tri-chambers (S, soma (left); M, methocel (middle); N, neurite (right)). (B) Cell bodies in S at 24 h post RABV P-mCherry infection (10⁵ ffu) in N. White arrow indicates an infected cell body with axonal connection to N. White asterisks indicate etched grooves (scale bar = 500 μm). (C) Quantification of % infected cell bodies at 72 hpi when axons were infected with different numbers of viral particles (10⁰ ffu–10⁶ ffu). (D) Quantification of % infected cell bodies from 20 h to 168 h post axonal infection with 10⁵ ffu. Black dots in (C) and (D) represent individual tri-chambers (from three (C) and two (D) independent replicates). Horizontal lines and errors bars represent mean ± SD for each condition with **p = 0.01, ***p = 0.0006, ****p < 0.0001 using one-way ANOVA (ns = not significant). (E) Anti-mCherry western blot of protein lysate from S compartment cell bodies at 72 h post axonal infection (10⁵ ffu). Arrowhead indicates band for P-mCherry fusion protein (~60 kDa). Asterisk indicates expected size of unfused mCherry protein (~27 kDa), which is not present in infected cells.