Figure 3: Immunofluorescence and immunoprecipitation analysis of BMAL1’s influence on PML and CLOCK nuclear localization and interaction.
From: Interactive Organization of the Circadian Core Regulators PER2, BMAL1, CLOCK and PML
Immunofluorescence analysis of Bmal1−/− and WT MEF cells transfected with plasmids expressing (a) Flag-CLOCK (red) and PML (green). We observed more than 20 cells and pictures are representative images. (b) Left: western analysis of cytosol and nuclear fractions for wild type and Bmal1−/− cells using the respective antibodies for PML, CLOCK and BMAL1. Right: statistical analysis of cytosol (C)/nuclear fraction (N) ratio. WT MEF cells were set to 1.0. Bars: SEM. P < 0.05. N.S.: Not significant. n = 4. (c) Immunofluorescence analysis of Bmal1−/− (upper) and WT MEF (lower) cells using the appropriate antibodies for PER2 and PML. We observed more than 20 cells and pictures are representative images. (d) Western analysis of IP from Bmal1−/− cells transfected with the indicated plasmid and stained with the indicated antibodies as stated (n = 2).
