Figure 3

Analysis of Pb sp and Pb SP expression during nitrogen starvation and during infection in murine macrophages. A: Western blot assay using the polyclonal antibody anti-Pb SP of protein extracts of. 1: yeast cells cultured in MMcM medium; 2: yeast cells cultured in the same medium deprived of nitrogen; 3: culture supernatant of yeast cells in MMcM medium; 4: the same as in 3 in the absence of nitrogen. B: Pb sp quantification by Real Time PCR. RNAs obtained were used to obtain cDNAs used to perform Pb sp quantification. Reactions were performed in triplicate and normalized by using α-tubulin expression. 1: Pb sp relative quantification in yeast cells incubated in MMcM medium for 4 h; 2: Pb sp relative quantification in yeast cells incubated in MMcM medium without nitrogen sources for 4 h; 3: Pb sp relative quantification in yeast cells incubated in MMcM medium for 8 h; 4: Pb sp relative quantification in yeast cells incubated in MMcM medium without nitrogen sources for 8 h. C: Pb sp quantification by Real Time PCR. 1: Pb sp relative quantification in mycelium. 2: Pb sp relative quantification in yeast cells. 3: Pb sp relative quantification in yeast cells during infection in macrophages. Asterisk denotes values statistically different from control (P ≤ 0.05).