Rodriguez, et al (2006): Figure 3

Supplementary Material From Singer Lab Publications:

Visualization of mRNA translation in living cells

J Cell Biol 175(1):67-76 (2006 October 9)

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Figure 3. Disrupting polysome structure with puromycin prevents the detection of ReAsH sites. (A) Overlay image of cells stained with ReAsH in the presence of 100 µg/ml cycloheximide showing GFP–ß-actin (green) and ReAsH staining (red) with a schematic representation of the time course of the procedure. Notice the presence of numerous ReAsH puncta throughout the cytoplasm. (B) Overlay image of a cell pretreated with 200 µg/ml puromycin stained with ReAsH in the presence of 100 µg/ml cycloheximide showing GFP–ß-actin (green) and ReAsH staining (red) with a schematic representation of the time course of the procedure. Notice the absence of any ReAsH puncta caused by the pretreatment with puromycin. (C) Graph of the relative number of ReAsH puncta per cell in cells treated with and without puromycin where the data are normalized to the number of puncta in the untreated sample. The disruption of polysomes leads to an approximately sevenfold reduction in the number of ReAsH puncta per cell as compared with control cells (P = 0.005). Untreated population, n = 5; puromycin-treated population, n = 8. Error bars are ± SEM.