Figure 3.
Early expression of RocG during sporulation is crucial for germination
(A) Spores of LR33 (Parental), LR38 (ΔrocR ΔahrC), LR134 (ΔrocR ΔahrC, PsspE-rocG) strains, lacking gudB, were disrupted for protein extraction. Equal amounts of protein extracts were subjected to Western blot analysis with antibody against RocG. SigA was monitored in parallel as a loading control using anti-SigA antibody.
(B) Spores of LR33 (parental), LR38 (ΔrocR ΔahrC), LR134 (ΔrocR ΔahrC, PsspE-rocG) strains, lacking gudB, were induced to germinate on an agarose pad supplemented with L-Ala (10 mM) (t = 0) and followed by time-lapse microscopy. Shown are phase contrast images captured at the indicated time points from a representative experiment out of three independent biological repeats. Scale bar, 1 μm.
(C) Quantification of the experiment described in (B). Data are presented as percentages of the initial number of the phase bright spores. Shown are average values and SD obtained from three independent biological repeats (n ≥ 300 for each strain).
(D) Spores of LR33 (Parental), LR38 (ΔrocR ΔahrC), LR133 (ΔrocR ΔahrC, PspoIID-rocG) strains, lacking gudB, were induced to germinate on an agarose pad supplemented with L-Ala (10 mM) (t = 0) and followed by time-lapse microscopy. Shown are phase contrast images captured at the indicated time points from a representative experiment, out of three independent biological repeats. Scale bar, 1 μm.
(E) Quantification of the experiment described in (D). Data are presented as percentages of the initial number of the phase bright spores. Shown are average values and SD obtained from three independent biological repeats (n ≥ 300 for each strain).