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Chikungunya virus dissemination from the midgut of Aedes aegypti is associated with temporal basal lamina degradation during bloodmeal digestion

Fig 2

Midgut BL associated collagen IV is diminished during bloodmeal digestion, which coincides with increased collagenase activity in the midgut between 12–36 h pbm.

(A) Detection of collagen IV in midguts of mosquitoes at different time points post-bloodmeal (pbm) and in sugarfed mosquitoes (Sugar) by Western blot using polyclonal antibodies generated against human collagen IV; two α chains of collagen IV (Col IVα) are detected. β-actin was detected as a loading control. Standard molecular sizes are indicated. (B) SDS-PAGE for the detection of human placenta collagen I, which was incubated with lysates prepared from mosquito midguts at different time points pbm. (C) In vitro activity assay to detect collagenase IV activity among lysates prepared from mosquito midguts at different time points pbm. Mean values with standard deviations from three independent experiments are shown. Different letters indicate significant differences based on one-way analysis of variance (ANOVA) followed by Tukey's multiple comparisons test (* at p ≤ 0.05). (D) Detection of collagen IV by Western blot in midguts of mosquitoes that had received a non-infectious or a CHIKV-containing bloodmeal at 1 and 2 days pbm/pi using polyclonal antibodies generated against human collagen IV. Images shown in panels A, B, D, are representative examples of repeated experiments. (E) Images of Ae. aegypti HWE females at 1 day pbm. The female on the left is fed to repletion ("full"), the one in the center is partially-fed ("half"), and the one on the right is sugarfed (no meal has entered the midgut). Detection of collagen IV in midguts of females, which were bloodfed to repletion ("full") or partially-fed ("half") at 1 and 2 days pbm by Western blot using polyclonal antibodies against human collagen IV; two α chains of collagen IV (Col IVα) were detected. β-actin was detected as a loading control.

Fig 2

doi: https://doi.org/10.1371/journal.pntd.0005976.g002