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. 2010 Jun 29;59(10):1355–1362. doi: 10.1136/gut.2010.207456

Figure 3.

Figure 3

Gp96 expression supports LF82 invasion. (A) Effect of pretreatment of Intestine-407 epithelial cells with anti-Gp96 antibodies on the invasive level of LF82. Intestine-407 cells were pretreated with rabbit polyclonal antibodies raised against Gp96 (Gp96 Ab) or with rabbit polyclonal antibodies (isotype control) diluted 1:200 or 1:500 for 30 min and then infected by LF82 bacteria. Invasion was determined after a 3 h infection period and after gentamicin treatment for an additional hour. Results are expressed as intracellular bacteria relative to those obtained for strain LF82 on non-treated cells (NT), taken as 100%. Each value is the mean±SEM of at least four separate experiments. *p<0.05 compared with the wild-type strain on untreated cells. (B) Western blot analysis of whole protein extracts from Intestine-407 cells using anti-Gp96 and anti-β-actin antibodies. Intestine-407 cells were non-transfected (NT), or transfected with 10 ng of small interfering RNA blocking Gp96 (gp96 siRNA), or non-specific siRNA as control. (C) Effect of gp96 siRNA on the invasive level of the wild-type strain LF82. Invasive bacteria were quantified as described in A. **p<0.01 compared with the wild-type strain on untreated cells.