Fig. 2: Growth kinetics and fluorescent microscopy of M. amalyticus and GAMs.
a Growth curve of G. amarae CON44T, GAM1, GAM2 and GAM6 in monoculture and coculture with M. amalyticus (MOI = 1.0) over 72 h. All GAMs exhibited lower final OD600 in comparison to the WT. As expected, growth in WT cocultures were heavily impaired, however each GAM tested displayed no reduction in growth when cocultured. b Direct dPCR measuring starting and final DNA copy number mL-1 of M. amalyticus in coculture. Abundance of M. amalyticus in GAM coculture was comparable to M. amalyticus monoculture. As expected, the copy number of M. amalyticus in G. amarae CON44T coculture displayed statistically significant increase in abundance (two-way ANOVA Å Ãdák’s multiple comparisons test, P < 0.0001). Black circles represent each data point (error bars representing standard error of the mean, n = 3 biological replicates) with white and grey bars representing starting (0 h) and end point (72 h) respectively. c Fluorescent microscopy of membrane-stained G. amarae CON44T-M. amalyticus coculture d and GAM7- M. amalyticus coculture, showing host and M. amalyticus cells in green and red, respectively. The white boxes represent the area shown in the zoomed micrographs to the right of each image. Due to reluctant uptake of membrane dye in GAM7, fluorescent excitation of G. amarae CON44T and GAM7 were 488 nm and 408 nm wavelengths, respectively. Images were captured every three seconds using a Nikon TiE inverted fluorescence microscope. Original magnification is 60x with scale bar representing 20 μm (see supplementary movies 2 and 3 for more detail). These results were independently reproduced three times.
