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. 2021 Mar 26;12:1907. doi: 10.1038/s41467-021-22169-1

Fig. 4. Characterization of Bp mutant BPSS0015.

Fig. 4

a BPSS0015 is upregulated at the protrusion stage during infection. First three boxes of each infection stage represent three biological replicates, the fourth represents the mean. b Number of plaques formed (PFU) by the BPSS0015 mutant is similar to wild-type Bp, indicating no defects in host cell invasion; however, reduced plaque diameters were observed, suggesting defects during the progression of infection (n = 3). c Rapamycin-induced (RAP) autophagy reduced the survival of both wild-type Bp and the BPSS0015 mutant (n = 3), whereas the inhibition of autophagy with 3-methyladenine (3-MA) significantly restored intracellular replication of the BPSS0015 mutant to wild-type level (n = 6). d Representative TEM images of macrophages infected with the BPSS0015 mutant show each bacterium surrounded by membrane structures. Scale bars = 1 μm. e Infected HEK293T cells stably expressing GFP-LC3 show co-localization of the autophagy marker, LC3 (green), with the BPSS0015 mutant (red), indicated by the white arrow, and no co-localization with wild-type Bp (red). Scale bars = 10 μm. f The BPSS0015 mutant is completely attenuated in BALB/c mice (n = 5) but bacterial burdens of surviving mice suggest that this mutant persists in vivo. Data in bar graphs represent means ± s.e.m. and analyzed via two-sided unpaired t-test. P values presented above relevant comparisons.