Fig. 5

Roles of autophagy-related proteins in the host response to Salmonella infection. (a) Region of interest (blue square) for confocal image acquisition of infected phagocytes in the blood circulation in d, h, l and p. (b, f, j and n) Survival curves of systemic S. Typhimurium infection experiments of atg5 (b), atg13 (f), rubcn kd(1) (j) and rubcn kd(2) (n) morpholino knockdown groups with their respective control morpholino-injected groups. Kd1 and kd2 refer to use of translation blocking (MO1-rubcn) and splice blocking (MO2-rubcn) morpholinos, respectively. For each comparison one representative of three replicates is shown. Survival was scored at 24, 48 and 72 hpi, following infection at 48 hpf. (c, g, k and o) Representative CFU counts of the S. Typhimurium infections of atg5 (c), atg13 (g), rubcn kd(1) (k) and rubcn kd(2) (o) morpholino knockdown groups with their respective controls at 24 hpi. Five embryos/larvae per time point were used and the log transformed CFU data are shown with the geometric mean per time point. (d, h, l and p) Representative confocal images of atg5 (d’), atg13 (h’), rubcn kd(1) (l’) and rubcn kd(2)’ (p’) morpholino knockdown embryos with their respective controls (d, h, l and p) at 4 hpi. (e, i, m and q): Quantification of GFP-Lc3-Salmonella associations in atg5 (e), atg13 (i), rubcn kd(1) (m) and rubcn kd(2) (q) morpholino knockdown groups with their respective controls at 4 hpi. Numbers of infected phagocytes positive or negative for GFP-Lc3-Salmonella associations were counted from confocal images and the percentages of Lc3-positive over the total were averaged from five embryos per group. Error bars represent the SD. Scale bars: 10 μm. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05, ns = non-significant.