Fig 1.
Percentage Viabilty of M. agalactiae-infected HeLa cells.
Viable HeLa cells were measured using Trypan blue staining at 24, 48 and 72 h p.i. Mean values ± SD from three independent experiments performed in duplicates are depicted.
Fig 2.
Phase contrast microscopic images showing the morphological changes in M. agalactiae-infected cells.
Images were taken at 24, 48 and 72 h p.i. at 10x magnification. Pointed arrows show the profound cell elongation and membrane blebbing in infected HeLa cells.
Fig 3.
DNA fluorescence staining showing chromatin condensation in M. agalactiae-infected HeLa cells 48 h p.i.
Cells were permeabilized and stained with DAPI, and images were taken using a fluorescence microscope. As a positive control, HeLa cells were treated with 1 μM staurosporine for 3 h and stained with DAPI. Bars, 10 μm.
Fig 4.
Caspase-3 cleavage in M. agalactiae-infected HeLa cells.
The graph represents the average relative density of cleaved caspase-3 from the immunoblots of M. agalactiae-infected (+ MA) and uninfected (- MA) HeLa cells at 24 and 48 h. As a positive control, HeLa cells were treated with 1 μM staurosporine (Sp) for 3 h. Experiments were performed at least three times, and average relative density values ± standard deviation are represented here. ‘*’ indicates p < 0.05.
Fig 5.
Percentage of cell lysis in M. agalactiae-infected HeLa cells.
Both time- and dose- dependent increase (p<0.05) was observed at 36 and 48 h p.i. with all tested MOIs compared to uninfected cells in absence of gentamicin. A significant decrease in cell lysis was observed in gentamicin-treated cells (+G) compared to gentamicin-untreated cells (-G) at 48 h p.i. (‘*’ indicates p<0.05).