Additional file 17. Visualization of KYSE-450 cellular morphology using hemacolor staining and detection of early apoptotic and late apoptotic/necrotic cells using flow cytometry. (A-D) Hemacolor staining of KYSE-450 cells incubated (A) without F. gastrosuis for 48 h and (B-D) with 500 µg F. gastrosuis strain CDW1 incubated for (B) 36 h, (C) 48 h and (D) 72 h. Following morphologic features can be seen: plasma membrane blebbing (white arrow), cell swelling (white star), cytoplasmic vacuoles (black star). Original magnification x400, scale bar represents 10 µm. (E-M) Representative population plots displaying viable (green, Annexin-V-FITC negative, PI negative), early apoptotic (red, Annexin-V-FITC positive, PI negative), late apoptotic/necrotic (blue, Annexin-V-FITC positive, PI positive) cells and late necrotic (orange, Annexin-V-FITC negative, PI positive) of KYSE-450 cells incubated (E) without F. gastrosuis lysate for 72 h; (F-I) with 500 µg F. gastrosuis strain CDW1 for (F) 24 h; (G) 36 h; (H) 48 h and (...