Fig 6. Loss of Ywp1 from the cell wall enhances binding of anti-β-1,3-glucan.

(A) A set of Ywp1 knockout and restored strains [22] was grown for 18 hr in YPD, fixed with formaldehyde, and labeled with anti-β-1,3-glucan as in Fig 4. A composite of ten flow cytometric profiles is shown. The quantity of Ywp1 was zero in knockout strains 3L1 and 4L1; attempted restoration of YWP1 generated a set of strains with no Ywp1 (16r2), a trace of Ywp1 (7r1), approximately half the normal amount (16s1 and 16s2), and close to the full amount (16r1), as indicated. Wild type strain DAY185 is strain BWP17 that was restored to prototrophy [33]. For further comparison, strains 16r1 and 16r2 were also labeled with just the secondary antibody to quantify background levels of fluorescence (black lines). (B) Ywp1 knockout strain 4L1 was grown at 30°C in liquid BMM13 culture; aliquots were taken at 4–73 hr after 1/20 dilution of a stationary phase culture into fresh BMM13; cells were fixed with formaldehyde and labeled with anti-β-1,3-glucan as above.