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. Author manuscript; available in PMC: 2016 Jul 21.
Published in final edited form as: Mol Microbiol. 2016 Feb 10;100(3):425–441. doi: 10.1111/mmi.13326

Fig. 3. Msb2 regulates the response to high temperature through the CEK pathway.

Fig. 3

A. Immunoblot of P~Cek1 levels in Msb2 deletions. Strains were incubated in pre-warmed YNB media with 1.25% GlcNac at 30°C or 37°C for 3 h. Total cellular protein (20 µg) from cell lysates was examined by immunoblot analysis with α-phospho p42/44 MAPK antibodies or α-Cek1 antibodies as a control.

B. To determine Msb2 shedding, control (CAI4) or cells expressing HA-Msb2 and derivatives were grown in YNB + Glu or YNB + GlcNac media (pre-warmed to 30°C or 37°C) for 3 h. Cells were removed by centrifugation, and proteins in the supernatant were precipitated by TCA. Normalized protein (20 µg) was examined by slot blot using anti-HA antibodies.

C. Cells were grown in YNB + 2% Glu for 3 h at 37°C and 42°C and observed by microscopy at 20X magnification. Arrows, examples of hyphae at 42°C Bar, 10 microns.