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. 2017 Oct 16;13(10):e1006671. doi: 10.1371/journal.ppat.1006671

Fig 6. Src-mediated EGFR transactivation is not required for Akt activation triggered by T. gondii but mediates parasite-induced early STAT3 activation.

Fig 6

A, A549 cells were treated with or without FAK inhibitor or were transfected with control siRNA or Src siRNA followed by challenge with RH T. gondii. Expression of total Akt and phospho-S473 Akt were examined by immunoblot. A vertical line was inserted between densitometry data of lysates from cells treated with control or FAK inhibitor, or transfected with control or Src siRNA to indicate that relative densities of phospho-Akt from infected cells were compared to bands from their respective uninfected (control) cells. Relative density of phospho-Akt for uninfected samples was given a value of 1. B, Human RPE cells were challenged with RH (type I) or ME49 (type II) strains of T. gondii. Expression of total STAT3 and phospho-Y705 STAT3 were examined by immunoblot. Relative densities of phospho-STAT3 in lysates from infected cells were compared to those from their respective uninfected (control) cells. Relative density of phospho-STAT3 for uninfected samples was given a value of 1. C, BV-2 cells were challenged with RH T. gondii followed by assessment of total STAT3 and phospho-Y705 STAT3 by immunoblot. D, A549 cells were challenged with either WT or Δrop16 T. gondii. Cell lysates were probed for total STAT3 and phospho-Y705 STAT3. Band densities in lysates from infected cells were compared to those from uninfected (control) cells. Densitometries for control bands were given a value of 1. E, A549 cells transfected with control siRNA or Src siRNA were challenged with RH T. gondii. Immunoblot and densitometries were assessed as above. F, A549 cells transfected with plasmid encoding either WT-EGFR or Y845F-EGFR were infected with T. gondii 48 h after transfection. Immunoblot and densitometries were assessed as above. G, NMuMG that express WT EGFR or EGFR AA mutant were challenged with WT or Δrop16 T. gondii. Immunoblot and densitometries were assessed as above. Densitometry data represent means ± SEM of 3 independent experiments.