Confocal microscopy of <i>M. hominis</i> invasion into, and bleb formation for evasion of HeLa cells.

<p>A, HeLa cells analysed at 48 h post infection with <i>M. hominis</i> at an MOI of 500 by confocal laser microscopy. Actin filaments were stained green (Alexa 488 phalloidin), nuclei blue (DAPI), surface-attached mycoplasmas magenta (mAb BG11 without fixation) and internalized mycoplasma cells red (mAb BG11 after fixation and permeabilisation). The four-coloured image (A1) is also shown in four single coloured pictures (A2) for a better discrimination of intra- and extra-cellular mycoplasma cells and demonstration of actin rearrangement at the location of intracellular mycoplasmas. B, HeLa cells chronically infected with <i>M. hominis</i>. Actin filaments are stained green (Alexa 488 phalloidin), nuclei blue (DAPI), <i>M. hominis</i> cells red (mAb BG11). A <i>M. hominis</i>-filled protrusion of the HeLa cell membrane bordered by actin (green) is marked by an arrowhead. Inset: SEM of a chronically infected HeLa cell with a prominent protrusion. C, HeLa cells chronically infected with <i>M. hominis</i>. LAMP3 proteins are stained green (mAb MEM-259), <i>M. hominis</i> cells red (mAb BG11), membranes cyan (Alexa Fluor 594 Wheat Germ Agglutinin) and nuclei blue (DAPI). Co-localisation of LAMP3 and <i>M. hominis</i> appears yellow in a three colour overlay (omitting stain of the membranes).