Figure 5

BEAS-2B cells induce chemokine gene expression in response to stimulation with acapsular C. neoformans. (A) BEAS-2B cells were left unstimulated (NS) (left panel) or stimulated (right panel) with acapsular C. neoformans MOI = 20 cultured at 37°C. Twenty-four hours later, RNA was extracted and analyzed by microarray for expression of inflammatory cytokines and their receptors. A box indicates the location of endogenous control (housekeeping) genes; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-2 microglobulin (B2M) were used for normalization. (B) Summary of relative gene expression following stimulation with C. neoformans. Data shown is representative of three independent experiments. *Not determined due to undetectable IL-8 expression in untreated cells. (C) Expression of CCL2, CXCL1, and CEBP/β was analyzed by RT-PCR; β-actin was used as an endogenous control. (D) Relative quantification of CXCL1, CCL2, and CEBP/β expression in untreated (white columns) and stimulated (black columns) BEAS-2B cells was determined by real time PCR. Results are representative of three independent experiments.