Kanamycin does not induce H1-T6SS-dependent secretion.
(A) Secretion assay performed on PAK cells incubated with increasing concentrations of kanamycin as indicated above the blot in µg/ml. Whole cell extracts are shown on the left of the blot, while supernatants are shown on the right. Blots have been probed with anti-PcrV (upper blot) and anti Hcp1 (lower blot). The expected location of each protein is indicated on the left. The PAKΔretS mutant strain, with a constitutively active H1-T6SS, is included as a positive control for secretion. * denotes that these samples have been diluted 1 in 10 with loading buffer to reduce the Hcp1 signal. (B) Western blot performed with anti-Tse3 antibodies on whole cell extracts prepared from PAK cells grown with increasing concentrations of kanamycin as indicated above the blots in µg/ml. PAKΔretS is included as control as it is known that Tse3 is induced in this background.