Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2009 Aug 19.

Published in final edited form as: Fungal Genet Biol. 2008 Jan 26;45(6):861–877. doi: 10.1016/j.fgb.2008.01.001

Fig. 6 — Filamentation on BSA and M199 agar plates (A). Overnight cultures of strains THE1-CIp10 and tetR-VPS1 were spotted onto BSA plates and incubated at 30 °C for 8 days. Robust filamentous structures emerging from the edge of each colony were observed in all strains, except in the tetR-VPS1 strain in the presence of doxycycline. Similar results were observed when these strains were spotted onto Medium 199 agar containing Earle’s salts, buffered with 150 mM HEPES to pH 7.5, and incubated at 37 °C for 5 days. Filamentation on Spider and 10% FCS agar plates (B). Overnight cultures were spotted onto either YPD containing 10% fetal calf serum (FCS) or Spider agar medium and incubated at 37 °C for 5 days. On Spider agar, robust filamentation extending above the surface of the plate can be seen in all strains except for the repressed tetR-VPS1 strain, which did not produce a raised colony. Similarly, on YPD agar with 10% FCS, filamentation was robust in all strains except in the tetR-VPS1 strain grown in the presence of doxycycline. Filamentation in liquid RPMI-1640 (C). Filamentation was assessed in RPMI-1640 medium buffered to pH 7.3 with 165 mM MOPS, with or without doxycycline. Medium was inoculated with 5 × 10⁶ cells/ml and incubated at 37 °C for 24 hr with constant shaking at 200 rpm. Strain THE1-CIp10 produced hyphae when grown in the presence or absence of doxycycline. In contrast, strain tetR-VPS1 produced masses of hyphae when grown under derepressing conditions, but not under repressing conditions. Filamentation in YPD containing 10% FCS (D). Pre-warmed medium was inoculated with 5 × 10⁶ cells/ml and incubated at 37 °C with constant shaking at 200 rpm. Induction of filamentation was followed by light microscopy at fixed time points from the time of inoculum, with a four hour time point shown. The tetR-VPS1 strain remained predominantly in the yeast form when grown in the presence of doxycycline, whereas, in contrast the other strains produced longer and greater numbers of filamentous structures.

Fig. 6 — Filamentation on BSA and M199 agar plates (A). Overnight cultures of strains THE1-CIp10 and tetR-VPS1 were spotted onto BSA plates and incubated at 30 °C for 8 days. Robust filamentous structures emerging from the edge of each colony were observed in all strains, except in the tetR-VPS1 strain in the presence of doxycycline. Similar results were observed when these strains were spotted onto Medium 199 agar containing Earle’s salts, buffered with 150 mM HEPES to pH 7.5, and incubated at 37 °C for 5 days. Filamentation on Spider and 10% FCS agar plates (B). Overnight cultures were spotted onto either YPD containing 10% fetal calf serum (FCS) or Spider agar medium and incubated at 37 °C for 5 days. On Spider agar, robust filamentation extending above the surface of the plate can be seen in all strains except for the repressed tetR-VPS1 strain, which did not produce a raised colony. Similarly, on YPD agar with 10% FCS, filamentation was robust in all strains except in the tetR-VPS1 strain grown in the presence of doxycycline. Filamentation in liquid RPMI-1640 (C). Filamentation was assessed in RPMI-1640 medium buffered to pH 7.3 with 165 mM MOPS, with or without doxycycline. Medium was inoculated with 5 × 10⁶ cells/ml and incubated at 37 °C for 24 hr with constant shaking at 200 rpm. Strain THE1-CIp10 produced hyphae when grown in the presence or absence of doxycycline. In contrast, strain tetR-VPS1 produced masses of hyphae when grown under derepressing conditions, but not under repressing conditions. Filamentation in YPD containing 10% FCS (D). Pre-warmed medium was inoculated with 5 × 10⁶ cells/ml and incubated at 37 °C with constant shaking at 200 rpm. Induction of filamentation was followed by light microscopy at fixed time points from the time of inoculum, with a four hour time point shown. The tetR-VPS1 strain remained predominantly in the yeast form when grown in the presence of doxycycline, whereas, in contrast the other strains produced longer and greater numbers of filamentous structures.