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Figure 7B shows a western blot of tagged substrate with and without induction of SspB. When SspB is induced/present in the cell, the tagged substrate is degraded rapidly by ClpXP.

The results of FACS (fluorescence activated cell sorting) is shown in figure 4. HeLa cells were treated with SubAB and by hour 20 it was evident that there was a definite increase in the number of cells in G0-G1 phase compared to time 0 as well as a decrease in the number of cells in the other stages. Figure 4 shows that SubAB causes cells to arrest in G1.

Figure 5 shows a western blot looking at the effect SubAB has on the levels of cyclin D1, cyclin D3, Cdk4, and Cdk6. This experiment was done to examine the underlying cause of cell cycle arrest. A significant downregulation of cyclin D1 occurred between 30 and 60 minutes after exposure to SubAB. Cyclin D is a G1 cyclin and the downregulation of cyclin D1 led to arrest in G1 of the cell cycle.

Figure 1A is a western blot that shows Vero cells incubated with and without SubAB. The antibody used was polyclonal goat anti-BiP. The antibody labeled a 72-kDa band in cells not incubated with SubAB which represents the intact BiP protein. In cells incubated with SubAB this band was diminished and a band at 28-kDa (the cleaved fragment) was visible. Only one cleavage fragment was visible because when the antibody was made, it was made against a peptide from the C-terminus of BiP so only the intact and C terminal portion of the protein would bind to the antibody and produce a band.

Figure 1B is a western blot which shows the cleavage of BiP by SubAB over a time span of 60 minutes. The antibody used was polyclonal goat anti-BiP. At 20 minutes the band at 28-kDa (part of the cleaved protein) became evident and by 30 minutes it was evident that the band at 72-kDa (the intact protein) had become diminished. Only one cleavage fragment was visible because when the antibody was made, it was made against a peptide from the C-terminus of BiP so only the intact and C terminal portion of the protein would bind to the antibody and produce a band.

The results from the two site-specific mutagenesis experiments indicate that flexibility between D149 and S193 is needed for better inducer binding.

Using immunocytochemistry and confocal analysis, figure 2 shows that VacA is localized to vacuoles, but not to mitochondria.

The western blot in figure 3 shows that no cytochrome c was released into the supernatant when isolated mitochondria were incubated with VacA.

Figure 4 (A-F) shows flow cytometric analysis using antibodies specific for active Bax to compare cells that were treated with VacA to cells that were not. Bax was activated when exposed to VacA.