Figure 2.
Localization of IHF and HU within an NTHI biofilm. Biofilm growth was initiated and maintained for 40 hr. Unfixed 40 hr biofilms were washed in PBS and incubated with (a) control IgG or (b) anti‐IHFA3, anti‐IHFB2 and anti‐HU 2 IgG and then incubated with goat anti‐mouse IgG conjugated to AlexaFluor® 405 and goat anti‐guinea pig IgG conjugated to AlexaFluor® 488 as required. NTHI were labeled with FilmTracer™ FM ® 4‐64. The biofilms were imaged using a x63 objective on a Zeiss 510 Meta‐laser scanning confocal microscope (Carl Zeiss, Thornwood, NY). NTHI are shown in gray, IHF in red and HU in green. Note that IHF and HU are found at distinct locations within the NTHI biofilm matrix.IHF, integration host factor; PBS, phosphate buffered saline; NTHI, nontypeable Haemophilus influenzae