Fig. 4

IBs processing is affected by the ΔcobB and ΔackA-pta mutations. The bacteria were grown and VP1GFP synthesis was induced as described in the legend to Fig. 1. At 3 h after IPTG induction (control), chloramphenicol (200 mg/L) was added to arrest protein synthesis and the cultures were incubated further at 28 °C. a After 1 and 3 h (+1 h, +3 h), cell samples were collected to isolate soluble fractions and IBs, which were subsequently analyzed by SDS-PAGE, immunodetection using anti-GFP antibodies, and densitometry. The level of VP1GFP after 3 h of induction (control) corresponds to 100%. b GFP fluorescence in soluble fractions and IBs was recorded in EnSpire plate reader (PerkinElmer). Specific fluorescence emission of VP1GFP was estimated as described in the “Methods” section. Error bars represent the standard deviation of three independent experiments