Figure 3.

Elongation of membrane wraps requires the activity of Src and PI3 kinases and Rho GTPases. Quantifications of the lengths of the segments of Lp filaments internalized in cells following the indicated treatments: (A) DMSO or Src inhibition with PP2, (B) DMSO or PI3K inhibition with Ly294002 (Ly), (C) DMSO or Cdc42 inhibition with ML141, (D) Water or Rac1 inhibition with NSC23766 (NSC), (E) Glycerol or Rho inhibition using C3 transferase (C3), (F) DMSO or Arp2/3 inhibition using CK666, (G) DMSO or mDia inhibition using SMIFH2, (H) DMSO or ROCK inhibition using Y27632 and (I) DMSO or Myosin II inhibition using blebbistatin. Cells were infected with RFP-Lp for 1h. Unbound bacteria were washed and the indicated inhibitors were added for 5 h before cells were fixed and external bacteria were immune-labeled. Internalization was quantified by measuring the lengths of the Lp segments that were inaccessible to the external antibody applied in non-permeabilized cells (see materials and methods). Data shown are mean ± SEMs from 3 independent experiments. Each segment of an Lp filament undergoing internalization was considered as an independent value. At least 100 filaments were analyzed in each condition. Statistical analyses were performed using Student's t-test, ^***p < 0.0001, ^**p < 0.001, ^*p < 0.05.