FIG 2.
The Sia signaling network impacts the surface adherence of planktonic cells through control of cell-associated Psl. (A) Adherence of matrix mutant strains to glass. Cells were incubated on a glass coverslip, rinsed and attached cells were immediately quantified by microscopy. (B) Representative immunoblot for Psl produced by sia mutants, extracted from midlog planktonic cells (OD600nm = 0.5). RNAP served as a loading control. (C) Quantification of relative Psl production calculated using blots in 2B. Psl band intensity was normalized to RNAP levels and then compared to wild-type (PAO1) cell-associated Psl. (D) Representative immunoblot for Psl from PAO1 and ΔsiaD throughout planktonic growth. Negative-control samples (ΔpslD) were processed at OD600nm = 2. At each time point 1 × 109 cells were processed. RNAP served as a loading control. (E) Growth curve of strains evaluated in 2D, in Lennox Broth. The time points collected for 2D are marked on the curve. (F) Quantification of relative cell-free and cell-associated Psl production calculated using blots in 2D. Psl band intensity was normalized to RNAP levels and then compared to PAO1 at OD600nm = 0.3. Presented as mean and standard deviation. N, 3 biological replicates, *, P < 0.05.