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. 2013 Jun 11;4(6):911–922. doi: 10.18632/oncotarget.1066

Figure 2. OHT-treatment induces ACD in MM cells.

Figure 2

A) Exponentially growing RPMI 8226 and LP-1 cells were treated with EtOH as vehicle or OHT at 10 μM for 24 h and stained with MDC. MDC-positive vesicles were observed with a confocal microscope (x400 magnification). B) Cells were treated with EtOH or OHT (10 μM) for 24 h and analyzed by transmission electron microscopy. C) RPMI 8226 and LP-1 cells were treated for 6 h with 10 μM OHT or vehicle, then subjected to immunocytochemistry with AbII alone as control or anti-LC3 Ab. Slides were analyzed by confocal microscopy (x180 magnification). D) RPMI 8226 and LP-1 cells were treated with 10 μM OHT (or EtOH) for 48 h followed by BAF A1 for 4 h (+) or not (-). Whole cell lysates were then analyzed by Western blotting with anti-LC3 Ab. D) Degradation of long-time proteins was determined in RPMI 8226 and LP-1 treated with EtOH, 10 μM OHT, 40 μM PBPE or 10 μM RU 58668 for 18 h in the presence or in the absence of 10 mM 3-MA. Experiments were repeated at three times in duplicate with comparable results. The data presented here are the means ± SEM of all experiments. *, p<0.001.