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. 2015 Apr 24;10(4):e0120497. doi: 10.1371/journal.pone.0120497

Fig 6. Alpha-toxin stimulated the phosphorylation of phospholipase Cγ-1.

Fig 6

(A) A549 cells were incubated with 1.0 μg/mL wild-type alpha-toxin or 1.0 μg/mL H148G alpha-toxin at 37°C. Cell lysates were separated by SDS-PAGE and blotted with antibodies to phospho-PLCγ-1, phospho-PLCγ-2, and phospho-PLCβ-3. (B) Phosphorylated of PLCγ-1 (black circles), PLCγ-2 (white circles), and PLCβ-3 (white squares) in untreated cells was set to 1. Values represent the mean ± SE; n = 5. (C) A549 cells were incubated with 1.0 μg/mL wild-type or H148G alpha-toxin at 37°C for 60 min. The cells were fixed, permeabilized, and stained with phospho-PLCγ-1 antibody and Hoechst 33342. Phospho-PLCγ-1 (red) and nuclei (blue) were visualized by fluorescence microscopy. Scale bar, 10 μm. (D) Phospho- PLCγ-1 fluorescence intensity was measured as described in Materials and Methods. Values represent the mean ± SE; n = 5; *, p < 0.01.