FIGURE 6.
Kinesin-1 promotes host cell invasion by UPEC. Bladder cells were pre-treated with aurintricarboxylic acid (ATA, 50 μm) for 3 h or with sodium orthovanadate (Na3VO4, 100 μm) for 1 h prior to infection with UTI89. After 2 h in the continued presence of drugs (a) intracellular and (b) total cell-associated bacterial titers were calculated and are expressed relative to controls treated with only carrier. Quantification of (c) internalized and (d) total cell-associated bacteria following infection of KLC2-silenced bladder cells by UTI89. e, semi-quantitative RT-PCR showing KLC2 and GAPDH message levels in bladder cells 72 h after transfection with either control nonspecific siRNA or KLC2-specific siRNA. Data in graphs are expressed relative to appropriate controls as the means ± S.E. of at least three independent experiments carried out in triplicate. f, 5637 bladder cells, following transfection with pKLC2_FLAG, were treated with either DMSO alone (D) or 300 nm TSA (T) for 3 h prior to lysis. The Western blot shows levels of FLAG-tagged KLC2, α-tubulin, acetylated tubulin, and actin present in total cell lysates (input) or in supernatants (Sup) recovered after polymerizing and spinning out microtubules (pellets). Similar results were obtained using hemagglutinin-tagged KLC2.