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. 2016 Jul 13;6:29521. doi: 10.1038/srep29521

Figure 5.

Figure 5

Clusterin prevents apoptosis by inhibiting Bax translocation to mitochondria (A) sCLU immunoprecipitated from Ly6C+ MDSCs treated or untreated 12 h with 20 uM curcumin (CUR) or 10 ng/ml DTX were subjected to western blot analysis with 6A7 anti-active Bax antibody. sCLU did not bind active Bax in control DMSO-treated Ly6C+ cells but did so after curcumin or DTX treatment. (B) RAW264.7 macrophages were transfected with antisense-scramble or CLU for 24 h and then treated with DMSO, 20 μM curcumin or 10 ng/ml DTX for 2 h. Cells were incubated with 125 nM MitoTracker Red CMXRos for 30 min at 37 °C (red fluorescence), then stained with anti-6A7 followed by Alexa Fluor 488-conjugated antibody (green fluorescence). Cells were examined by confocal microscopy (magnification, ×630). Active Bax shows little translocation to mitochondria in scrambled control but almost complete colocalization in anti-sense CLU-transfected macrophages upon curcumin or DTX treatment. One representative set of images of three independent experiments is shown.