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. 2012 Feb 20;196(4):497–511. doi: 10.1083/jcb.201108118

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© 2012 Bakkar et al.

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Figure 2. — The alternative NF-κB subunit RelB regulates mitochondria and a glycolytic phenotype. (A) RNA was extracted from TA or soleus of 5-wk-old mice (n = 4 each), and mitochondrial genes were quantitated by real-time PCR. Asterisks denote significance for COX IVi in TA (P = 0.04) and soleus (P = 0.0006) and COX Va in TA (P = 0.03) and soleus (P = 0.003). (B) Microarray analysis from soleus of adult RelB^+/+ and RelB^−/− littermates. Mitochondrial genes that are most down-regulated are shown. Green and red colors indicate lower and higher RNA expression, respectively. (C) Gene ontology classification of down-regulated genes in RelB^−/− soleus muscles (n = 3). Shown in parentheses is the number of genes that changes in each category. (D) Representative soleus from RelB^+/+ and RelB^−/− mice. (E) Total DNA was extracted from hind-limb muscles of 5-wk-old RelB^+/+ and RelB^−/− mice (n = 3 each), and relative mitochondrial to nuclear copy numbers were calculated for TA (P = 0.04), soleus (P = 0.001), and gastrocnemius (Gas; P = 0.02). (F) TA muscles from 4-wk-old mice were harvested and stained for SDH. Bar, 200 µm. (G) RNA was prepared from soleus of RelB^+/+ and RelB^−/− adult mice as in A, and levels of myosin isotypes were determined by real-time RT-PCR. Asterisk denotes significance for MyHC IIa (P = 0.02) and MyHC IIb (P = 0.01). (H) TA homogenates from RelB^+/+ and RelB^−/− mice (n = 4) were prepared and measured for lactate normalized to micrograms of muscle proteins. Asterisk denotes significance. (I) 4-wk-old RelB^+/+ and RelB^−/− mice were housed in individual metabolic cages for 72 h, and RER, total activity, and food intake were measured. Results shown are the mean of four mice per genotype, monitored during light and dark hours. *, P < 0.05. Error bars are means ± SEM.