The Glyceraldehyde-3-Phosphate Dehydrogenase and the Small GTPase Rab 2 Are Crucial for Brucella Replication (original) (raw)
Figure 5
The inhibition of GAPDH expression and the dominant negative of Rab 2 affects B. abortus replication at 48 h p.i.
(A) Fold increase of intracellular CFUs at 48 h p.i. of Hela cells infected with B. abortus Ds Red. Infected cells were transfected or not with GFP, myc Rab 2, myc Rab 2 I119, GFP Rab 1 or GFP Rab 1 S25N at 2 h p.i. Data are means±SD of three independent experiments. (B) Confocal micrographs of Hela cells infected with B. abortus Ds red and transfected with myc Rab 2 or its dominant negative myc Rab 2 I119. The myc tag was immunostained with anti-myc antibody (Scale bars: 10 µm). (C, D, E) Quantification of the percentage of BCV positive for LAMP-1 (C), Calnexin (D) or Cathepsin D (E) in myc Rab 2 I119 transfected HeLa cells (open circle) or not (open square) and infected with B. abortus wild-type. (F) Western blotting of cellular extracts from cells transfected with GAPDH siRNA or siRNA control. Immunoblotting with anti-GAPDH and anti-actin antibodies. (G) Fold increase of intracellular CFUs at 48 h p.i. of transfected Hela cells with GAPDH siRNA or siRNA control and infected with B. abortus. Data are means±SD of three independent experiments. (H) Confocal micrographs of Hela cells infected with B. abortus Ds red and transfected with GAPDH siRNA or siRNA control. GAPDH was immunostained with anti-GAPDH antibody (Scale bars: 10 µm).