Colocalization of rNadA with HSP90.

A: Chang cells, stained in vivo with a rabbit polyclonal anti-HSP90, were incubated with rNadA for 1 h at 37°C, then fixed, permeabilized and double stained for rNadA (green) and HSP90 (red). Panels are taken from two different experiments. Merged images are also shown. Scale bar 10 µm. B: Quantification of rNadA and Hsp90 colocalization. rNadA column indicate the percentage of rNadA immunofluorescent pixels colocalizing with HSP90 immunofluorescent pixels. Conversely, HSP90 columns indicate the percentage of HSP90 immunofluorescent pixels colocalizing with rNadA immunofluorescent pixels. Data are mean ± s.e.m representative of two independent experiments, each assessing 20–25 cells. C: Chang cells pre-treated overnight with either vehicle (upper panel) or 0.5 µM 17-AAG (lower panel), were incubated with 200 µg/ml rNadA for 1 h, 4 h and 16 hrs as indicated, then fixed, permeabilized and stained for rNadA (red). Intracellular accumulation of rNadA clusters are indicated by arrows. Scale bar 10 µm. D: Quantification of IF intensity of rNadA in the experiment illustrated in panel C. Data are mean ± s.e.m representative of two independent experiments, each assessing 20–25 cells, and expressed as Arbitrary Units (A.U.). ***p