Identification of App1 as a regulator of phagocytosis and virulence of Cryptococcus neoformans
J. Clin. Invest. Chiara Luberto, et al. 112:1080 doi:10.1172/JCI18309 [
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Figure 3Disruption of
APP1 gene. (
a) Schematic representation for disruption and reconstitution of
APP1 gene. (
b) Southern analysis. Genomic DNA from
C. neoformans WT H99,
Δapp1, and
Δapp1Rec strains were extracted, digested with
PstI, electrophoresed in a 0.7% agarose gel, transferred to a Nytran membrane, and blotted against 5′-UTR and
ADE2 probes. The
Δapp1 strain showed a deletion of the
APP1 gene with insertion of the
ADE2 gene. Reintroduction of the
APP1 gene in the
APP1 locus is shown in the
Δapp1Rec strain. (
c) Western analysis of cell lysate from WT,
Δapp1, and
Δapp1Rec cell cultures using rabbit anti-App1 polyclonal antibodies. App1 protein is absent in the
Δapp1 mutant and reconstituted in the
Δapp1Rec strain. (
d) Western analysis of crude cytoplasmic extract, serum of two AIDSpatients, serum from two normal subjects, and growth culture medium. App1 is secreted extracellularly in vitro and during cryptococcal meningoencephalitis. HYG, hygromycin B gene.
