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. 2012 Jul 27;21(21):4680–4702. doi: 10.1093/hmg/dds309

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© The Author 2012. Published by Oxford University Press

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — QC autophagic response is impaired in mtDNA-depleted cells. (A) LC3B immunostaining (green) of Rho0 cells maintained in serum (+) or serum (−) conditions and NL treatment, following growth in the presence [pyr/urd (+/+)] or absence [pyr/urd (−/−)] of pyruvate and uridine. Hoechst 33342-stained nuclei are in blue. (B) Mean number of LC3B-positive vesicles per cell profile [n= 3, **P< 0.01 versus S+ pyr/urd (+/+); ^###P< 0.001 versus S− pyr/urd (+/+)]. Scale bar: 10 µm. (C) Immunoblot for endogenous LC3B from Rho0 cells maintained in serum (+) or serum (−) and NL treatment following growth in pyr/urd (+/+) or pyr/urd (−/−) conditions. (D) Densitometric analysis of LC3B endogenous levels [n= 5, *P< 0.05, **P< 0.01, versus S+ pyr/urd (+/+); ^##P< 0.01, versus S+ pyr/urd (−/−); ^&&P< 0.01, versus S− pyr/urd (+/+); ^$$P< 0.01, versus S− pyr/urd (−/−)]. (E) Determination of autophagic vacuole (AV) levels [n= 5, *P< 0.05, versus S+ pyr/urd (+/+)]. (F) Assessment of autophagic flux (n= 5).