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. 2010 Mar 5;6(3):e1000792. doi: 10.1371/journal.ppat.1000792

Figure 1. Anthrax toxin enter cells by clathrin mediated endocytosis.

Figure 1

A: Hela cells were transfected 72 hrs with control siRNAs or siRNAs against Clathrin Heavy Chain (CHC), the dynamin 2 (Dyn-2) or Epidermal growth factor receptor pathway substrate 15 (Eps 15). Cells were incubated with 500 ng/ml PA63 for 1 hr at 4°C and different times at 37°C and cell extracts (40 µg of proteins) were analyzed by SDS-PAGE and western blotting to reveal PA monomer and SDS-resistant heptamer (pPA7mer). B: Cell extracts (40 µg of protein) from Raw, BHK and Hela–transfected or not with mouse CMG2-4–cells were analyzed by SDS-PAGE and Western blotting for the expression of CMG2. C: Stable BHK21-tTA/anti-CHC cells maintained in tetracycline (control cells) or not (CHC AS) were incubated with 500 ng/ml PA63 for 1 hr at 4°C and different times at 37°C and extracts (40 µg of proteins) were analyzed by SDS-PAGE and western blotting to reveal PA63 and SDS-resistant heptamer (pPA7mer). DE: Hela cells were transfected 72 hrs with human CMG2-V5 and with control siRNAs or siRNAs against CHC. Cells were then treated as in A. Cells extracts (40 µg of proteins) were analyzed by SDS-PAGE and western blotting to reveal the different forms of PA, CMG2-V5 and the N-terminus of the LF target MEK1 (MEK1 (N)).