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. Author manuscript; available in PMC: 2018 Mar 19.
Published in final edited form as: J Biol Rhythms. 2017 Dec 27;33(1):99–105. doi: 10.1177/0748730417745178

Figure 2.

Figure 2

Circadian gating of Aspergillus fumigatus clearance from the mouse lung. (A) Flow cytometric analysis of Dectin-1 cell surface expression (BioLegend, #144303) on BMDMs, gated on CD11b+ cells (n=1). (B) Flow cytometric analysis of dsRed+ signal (A. fumigatus), gated on F4/80+ macrophages, after one-hour co-incubation with A. fumigatus in culture. Serum-synchronized BMDM from either wild-type (black) or mPer1m/mmPer2m/m (white) background were treated with swollen dsRed-expressing A. fumigatus Af293 conidia at a multiplicity of infection of one (n=1). Conidia were swollen by prior incubation in DMEM media at 37 °C with shaking for six hours. (C) Time course of intratracheal inoculation of 107 Af293 conidia into wild-type C57BL/6J (two-month-old male C57BL/6J mice from Jackson Laboratories were fed rodent chow ad libitum, maintained under constant environmental conditions, and entrained to a 12 hr light/dark cycle, light period from 7am to 7pm) or mPer1m/mmPer2m/m C57BL/6 immunocompetent mice (n=4 per group) and subsequent lung harvest after 16-hour incubation for A. fumigatus colony-forming unit (CFU) count on YPD (yeast extract, peptone, and dextrose) agar plates. (D) Intratracheal inoculation of 107 Af293 conidia into wild-type C57BL/6J mice (n=10 per group) at ZT0/ZT12 and lung harvest after 16-hour incubation for CFU count on YPD. (E) Intratracheal inoculation of 105 Af293 conidia into wild-type C57BL/6J mice (n=10 per group) at ZT0/ZT12 and lung harvest after 14-hour incubation for CFU count on YPD. Error bars represent 95% CI. *p<0.05 by Student’s two-tailed t-test.