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. Author manuscript; available in PMC: 2018 Sep 25.
Published in final edited form as: Curr Biol. 2017 Sep 14;27(18):2878–2886.e5. doi: 10.1016/j.cub.2017.08.032

Figure 2. Chs3-dependent chitin synthesis at the bud neck is increased in hof1Δ cells.

Figure 2

(A) hof1Δ cells are sensitive to Calcofluor White (CW). Strains YEF473A (wild-type, WT), YEF4600 (hof1Δ), YEF2368 (cyk3Δ), YEF4559 (chs3Δ), YEF2197 (chs4Δ), and YEF2769 (bni4Δ) (See also Table S1) were streaked out on an YPD plate and an YPD plate containing 25 μg/ml CW and incubated at 25°C for 3 days before documentation.

(B) Chitin level in the cell wall of hof1Δ cells is increased. Chitin levels in the total cell walls of the strains listed in (A), which were grown in YM-1 medium at 25°C or 37°C, were measured as described in STAR METHODS. Data are averaged from 9 and 4 independent experiments for the 25°C and 37°C samples, respectively. Error bars represent SEM (standard error of the mean).

(C–D) Chitin level at the bud neck of hof1Δ cells is increased from G2/M to telophase in a Chs3-dependent manner. Chitin at the bud neck of WT (YEF473A), hof1Δ (YEF4600), and hof1Δ chs3Δ (YEF2757) (See also Table S1) cells was visualized by CW staining (C) and then quantified as described in STAR METHODS (D). Cell cycle stages were estimated based on nuclear staining by DAPI. The number of cells used for quantification was: WT, n=34, 26, and 27; hof1Δ, n=12, 24, and 28; hof1Δ chs3Δ, n=10, 17, and 13 for G1/S, G2/M, and telophase, respectively. Error bars represent SEM (standard error of the mean).