Expression of hypha-associated Sap proteins during phagocytosis. (A) C. albicans SC5314 was cultured in YPD medium at 30°C to maintain yeast form, in RPMI-FBS (which contained 10% FBS and 0.4% glucose) medium at 37°C to generate hypha-form, and co-cultured C. albicans with THP-1 human monocytes in RPMI-FBS medium at 37°C. The expression of hypha-associated SAP genes was identified by reverse transcription-PCR analysis using gene specific primers. Y: YPD, S: RPMI-FBS, C: co-culture. EFB1 gene is an internal control. (B) Detection the secreted Sap5 protein in C. albicans and THP-1 co-culture condition by Western blot analysis. The * is a non-specific signal. Protein samples of the upper panel and the lower panel were manipulated simultaneously. (C) Detection the secreted Sap proteins on hyphae surfaces of C. albicans by immunofluorescence staining using polyclonal anti-Sap6 antibody as primary antibody. The polyclonal anti-Ach1 antibody is a negative control. (D) Detection the expression of secreted hypha-associated Sap proteins during phagocytosis by immunofluorescence staining. The majority of fluorescent signals distributed on the distal surface of hyphae.