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. Author manuscript; available in PMC: 2010 Jul 15.
Published in final edited form as: Exp Cell Res. 2009 Apr 14;315(12):2001–2011. doi: 10.1016/j.yexcr.2009.04.003

Figure 4. Shank3 deficient cells are less able to support efficient EPEC pedestal formation.

Figure 4

(A) HeLa cells were transfected with two levels of control shRNA vector (black bars) or one of two shank3-targeting shRNA vectors (grey and white bars). After 48 hours knockdown was assessed using real-time quantitative RT-PCR with Shank3 specific primers and normalized to GAPDH. Shown are means of two experiments, each using two independent cDNA samples, with standard deviations. A representative western blot following high-level (1000 ng) transfection of shRNA vectors is shown. Levels of endogenous Shank3 (as determined with anti-Shank3 antibody) are reduced in cells receiving Shank3-directed shRNAs 1 and 2, compared to control-transfected cells. Anti-tubulin is shown as a protein-level control.

(B) HeLa cells, WT MEFs or Nck−/− MEFs were transfected with shRNA constructs for 48 hours, then infected with EPEC for 6 hours as described. Pedestal formation was assessed by F-actin staining and is expressed as a percentage of total adherent bacteria. Means of data pooled from three independent experiments are shown, at least 40 cells per condition were counted in each experiment. Error bars indicate 95% confidence intervals, * signifies statistical significance p<0.05. Significance was measured using a two-sample T-test, comparing to the relevant shRNA control samples. Data is representative of three independent experiments.