FIG 1.

Microarray analysis reveals A3G-upregulated and -downregulated genes. (A) Box plots of log₂ values for quantile-quantile normalized probe signal intensities. Total RNA was isolated independently two times each from control Vero-023 cells and A3G-expressing Vero (Vero-A3G) cells and labeled. Bars 1 and 3, RNAs from Vero-023 control cells; bars 2 and 4, RNAs from Vero-A3G cells. The probes were hybridized to Gene Chip rhesus macaque genome arrays (Affymetrix) according to the manufacturer's instructions. (B) MA plot (intensity log₂ fold change [M] plotted against the average log2 intensity [A]) for the comparison between treated and untreated cells. Mean intensities of both groups (x axis) were plotted against log₂ fold change (y axis). Blue circles around dots show the first 15 genes with the lowest P values. (C) Numbers of all probe sets, of significantly (adjusted P values of <0.05) up- and downregulated probe sets, and of more than 2-fold-up- and downregulated probe sets. (D) List of the best up- and downregulated genes, with the alteration (ratio) of transcript expression and significance (adjusted P values) as determined by the four microarrays. (E) Total RNA was isolated from control (Vero-023) and A3G-expressing (Vero-A3G) cells. Four amounts of RNA (4, 2, 1, and 0.5 μg) were reverse transcribed into cDNA. Four microliters of each of cDNA product was amplified by PCR and analyzed on 1% agarose gels. (F) Protein lysates of Vero-023 and Vero-A3G cells were separated by 10% SDS-PAGE, blotted to nitrocellulose, incubated with antibodies to REDD1, KDELR2, MOSC2, ACY1, and PRDX2 (and GAPDH as a control), and visualized using the ECL system. Quantifications of the Western blots normalized to the value for Vero-023 cells are shown below each blot.