Figure S1F - Western blot analysis of RelA protein levels before and after starvation

Western blot analysis of RelA protein levels before and after starvation. MG1655, MG1655relA::HTF, MG1655relAΔRRM::HTF, MG1655relAΔZFDΔRRM::HTF, MG1655relAH432E::HTF where grown exponentially in MOPS minimal medium with 0.2% glucose at 30°C. Before starvation samples (1 mL) were collected at OD600 = 0.2 and the cultures starved for isoleucine by addition of L-Valine to a final concentration of 500 μg/mL. After 30 min of starvation, a second sample was collected. The samples were denatured and separated by SDS-PAGE, transferred to a PVDF membrane and the hexa Histidine tagged protein detected using pentahis antibodies.