Figure 5
SopD2 targets AnxA2 in vitro. (a) Representative blot of reciprocal pull-down of AnxA2 and SopD2 or PipB2. Concentrated culture supernatants of S. Typhimurium strains secreting SopD2-2HA, PipB2-2HA, or HA empty vector were mixed with His6-AnxA2. Pull-downs were analyzed by Western blot using α-HA or α-AnxA2. IB: Immunoblot. Leftmost lane: pull-down of SopD2-2HA in the absence of AnxA2. Lane second from the left: pull-down of SopD2-2HA in the presence of AnxA2. Middle lane: pull-down of PipB2-2HA in the absence of AnxA2. Lane second from the right: pull-down of PipB2-2HA in the presence of AnxA2. Rightmost lane: pull-down of empty vector expressing a HA-tag in the presence of AnxA2. Original Western blots are shown in Supplemental Fig. S4. (b) Fold change determined by densitometry of 3 Western blots from independent experiments. Mean fold change ± standard deviation is shown. Fold change = (Densitometry of effector + AnxA2)/(Densitometry of effector). Dashed line = 1.
