Fig. 3

Effects of nitric oxide (NO) generators on β chemokine production. (A) Analysis of chemokine β peptides in the supernatants of monocyte cultures. Monocyte cultures were stimulated with 0.5 ng/ml lipopolysaccharide (LPS) alone or in combination with NO donors, S-nitroso-N-acetyl-penicilamine (SNAP; 40 µM) or sodium nitroprusside (SNP; 100 µM). 18 hr after the stimulation, chemokine β peptides were assayed by ELISA. Data shown are mean ± SEM of the triplicate cultures, and ^* indicates p-value ≤ 0.05 calculated using the two-tailed t-test. (B) RNase protection analysis of human immunodeficiency virus-type 1 (HIV-1)-infected and NO-treated monocytes. HIV-1-infected and control monocyte cultures were treated with 0.5 ng/ml LPS. To some cultures, donors of NO were added together with LPS at the following concentrations: SNP at 100 µM, spermine bis(nitric oxide) adduct (NONOate) at 10 µM, SNAP at 500 µM. 2 hr after stimulation, cytoplasmic RNA was extracted and assayed by RNase protection method. Bands corresponding to protected fragments of MIP-1α, MIP-1β, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are marked. Intensity of the bands is directly proportional to the amount of corresponding RNA.