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. 2009 Dec 4;11(1):37–44. doi: 10.1038/embor.2009.242

Figure 4.

Figure 4

DOR loss-of-function reduces autophagy. (A,B) C2C12 myoblasts were previously infected with lentiviruses encoding scrambled RNA, DOR siRNA1 or DOR siRNA2. Endogenous LC3 was detected by immunofluorescence. LC3-positive puncta were counted in 100 cells per group under basal conditions or starved for 20 min. Scale bars, 10 μm. *Significant effects of starvation, P<0.001; τsignificant effects caused by DOR knockdown, P<0.001. (C,D) Scramble or DOR siRNA1 C2C12 cells were processed by transmission electron microscopy. Arrows indicate autophagosomes. Insets are autophagosomes. Scale bars, 1 μm. Autophagosomes were counted in 50 randomly chosen cells. *Significant effects of DOR repression, P<0.001. (E) Scramble or DOR siRNA1 C2C12 cells were incubated in DMEM with and without 10 mM 3MA. *Significant effects of DOR knockdown, P<0.01; τsignificant effects caused by 3MA, P<0.01. 3MA, 3-methyladenine; DMEM, Dulbecco's modified Eagle medium; DOR, diabetes- and obesity-regulated gene; LC3, microtubule-associated protein 1A/1B-light chain 3.