FIG 1.

Micrococcal nuclease digest of L. donovani nuclear DNA. A total of 5 × 10⁷ cells per sample were permeabilized and treated with MNase for 5 min at room temperature. The reaction was stopped, and samples were incubated for 2 h at 37°C with proteinase K. DNA was precipitated with ethanol, and 3 μg of DNA was analyzed on a 1.5% agarose gel. The intensity of mononucleosome-derived DNA bands was quantified using Image J software and normalized to control. (A) MNase dose testing and impact on the intensity of mononucleosome-derived DNA bands. (B) 40 units/ml MNase. (C to F) 80 units/ml MNase. (B) Treatment of L. donovani promastigotes with hydroxyurea for 24 h at 25°C. (C) Treatment of L. donovani promastigotes with radicicol (RAD) at various concentrations for 72 h at 25°C. (D) The same as in panel C, but with L. donovani (HSP90rr) promastigotes. (E) L. donovani promastigotes at 37°C for 72 h. (F) Axenic L. donovani amastigotes at day 4 of differentiation. Significance for panels E and F was tested using the ratio-paired t test. ****, P ≤ 0.0001; **, P ≤ 0.01; low temperature (LT) = 25°C; high temperature (HT) = 37°C.