FIG 5.

Induced E. faecalis cells that express early induced transcripts also express late induced transcripts. (A) E. faecalis OG1RF/pCF10 was harvested for HCR labeling at 30 min after addition of 2.5 ng · ml⁻¹ C. Green and red, transcripts labeled by HCR with Alexa Fluor 488 and Alexa Fluor 647, respectively; blue, nucleic acids (primarily DNA) labeled with Hoechst 33342 highlight individual cells. (Top) Induced prgB (green, early gene) and pcfC (red, late gene) HCR-labeled transcripts. (Bottom) Induced pcfG (green, late gene) and Q_L (red, early gene) HCR-labeled transcripts. Bars, 10 μm. (B) Schematic showing the distances between the genes whose transcripts were probed using HCR. See Fig. 1 for a more complete map. Not shown is a control experiment in which an identically induced aliquot of donor cells was subjected to HCR analysis for both Q_L and pcfG expression but where the fluorophores used for detection were swapped; the fractions of positive cells for each transcript were not detectably different from those shown in the images in the lower part of panel A.