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. 2015 Jun 11;10(6):e0130189. doi: 10.1371/journal.pone.0130189

Fig 6. Nef myristoylation and the acidic cluster are essential to induce iNOS.

Fig 6

BV-2 cells were left untreated or incubated with 100 ng/ml wild type (WT), G2A, two different preparations of the EEEE→AAAA mutant (4EA’ and 4EA”) and ΔN-terminal deleted myr+NefSF2 proteins. (A) iNOS mRNA was measured by real time RT-PCR using total RNA isolated from cells treated for 6 h. (B) Total cellular extracts obtained from cells treated for 24h were analyzed by Western Blot using anti-iNOS antibodies, whereas β-tubulin expression levels were used as an internal loading control. (C) Supernatants collected from cells treated as in (B) were measured for NO2 - content using the Griess colorimetric assay. One out of three independent experiments is shown.