FIG. 4.

Sorting and postsorting analysis of mixed cultures. Mixed cultures of M. smegmatis mc²155 expressing the SIINFEKL epitope (Ms-pMP10), producing white colonies, and M. smegmatis mc²155 (Ms-dsRed), producing red colonies, were stained with anti-SIINFEKL serum followed by goat anti-rabbit IgG-Alexa Fluor 647 (AF647) and goat anti-rabbit IgG-AF488. Bacteria with double-negative (AF488^lo AF647^lo) and double-positive (AF488^hi AF647^hi) staining were sorted and then plated onto 7H10 plates containing kanamycin. After growth, the percentage of red colonies in each plate was measured. “TNTC” indicates that colonies were too numerous to count. We performed sorting and plating experiments three times with the same results.