(A and B) Strategies to block translation in the forespore (A) and to
determine the degree to which IleS-ssrA* degradation reduces forespore
translation (B).
(C) Fluorescence microscopy of sporangia expressing
PspoIIQ–CFP without
IleS degradation (left) or with IleS degradation in the forespore (right).
Membranes were stained with FM4–64 (red). CFP images (blue) used
identical exposures and adjustments to directly compare fluorescence intensity.
Scale bar, 1 μm.
(D) Total CFP fluorescence in the forespore (x axis) versus forespore
volume (y axis) of sporangia without (blue) or with (red) IleS degradation in
the forespore. Each dot represents an individual forespore. Solid dots represent
the average CFP intensity and volume of wild-type (blue) and IleS-depleted
forespores (red). Error bars represent standard deviations. IleS degradation
significantly reduced CFP intensity (p < 0.0001) but has no effect on
forespore size (p = 4625) compared to the control.
(E) Timelapse fluorescence microscopy of wild-type (control) and after
IleS degradation in the forespore (IleSdeg.).
(F) Change in forespore volume during engulfment in wild-type (WT,
green) and after forespore degradation of IleS (IleSdeg., red). Error
bars represent standard deviation.
See also Figure
S4.