E significant increment of caspase-3 (a downstream apoptotic factor) activity (Fig. 3E). We also observed increased expression of macrophage markers (F4/80 and CD68) in SMS1KO WAT, probably suggesting enhanced invasion of macrophage (Fig. 3F).Hyperlipidemia and Lipodystrophy in SMS1-KO Mice Are Rescued by Anti-oxidant TreatmentOur results support the idea that WAT cells of SMS1-KO mice are functionally damaged by increased Arg8-vasopressin web buy 117793 oxidative stress. Indeed, triglyceride levels were greatly increased in blood plasma of SMS1KO compared to wild-type mice, indicative of hyperlipidemia (Fig. 6A, also see Fig. 2A). In 
particular, triglyceride levels of chylomicron (CM) and very-low-density lipoprotein (VLDL), which normally function in muscle or are stored in adipocytes [43], were greatly increased in blood plasma, suggesting that lipid storage function of adipose cells is compromised in SMS1-KO mice (Fig. 6A). To further examine whether oxidative stress mediates these phenotypes, we supplied the anti-oxidant N-acetyl cysteine (NAC) in the drinking water of mutant mice and analyzed potential changes in blood plasma triglyceride levels (Fig. 6A). NAC-treated SMS1-KO mice showed robust decreases in blood triglyceride levels, almost to levels comparable to wild-type mice. Finally, we subjected mice to fasting for 30 hours and assessed triglyceride levels in blood plasma (Fig. 6B). Those levels increased in fasting wild-type mice and in SMS1-KO mice supplied normal drinking water. However, CM levels in fasting SMS1-KO mice were significantly decreased compared to wild-type mice. By contrast, SMS1-KO mice supplied with NAC-containing drinking water exhibited partially recovered CM levels. The volume of SMS1-KO epiWAT was also partially rescued by NAC-treatment (Fig. 6C). Altogether, these results indicate that increased oxidative stress underlies a failure of lipid storage function in adipocytes of SMS1-KO mice.Oxidative Stress Response, the Mitochondrial Stress Response and Mitochondrial Biogenesis Are Enhanced in SMS1-KO WATTo confirm whether oxidative stress occurs in SMS1-KO WAT, we examined expression of mRNAs encoding ROS detoxification enzymes. In SMS1-KO WAT, expression of some isoforms of peroxiredoxin (Prdx), which reduces hydrogen peroxide and alkyl hydroperoxides, was increased (Fig. 4A, Table S1). Expression of catalase, which catalyzes decomposition of hydrogen peroxide, also increased. Other ROS detoxification enzymes, such as glutathione peroxidase (Gpx), 15755315 superoxide dismutase (SOD) and glutaredoxin (Glrx), also showed increased expression. These data indicate that the oxidative stress response is enhanced in SMS1KO WAT. Next we examined expression of mRNAs encoding mitochondrial stress response-related factors. Most of these, including mitochondrial chaperones (mtHsp70, mtHsp40) and mitochondrial proteases (LONP1, ClpP, YME1L1, AFG3L2), were upregulated in SMS1-KO WAT (Fig. 4B, Table S1). On the other hand, mRNA expression of calnexin, an ER chaperone, or HSP70-1, a cytosolic chaperone, was not affected. We also observed increases in mRNAs encoding factors relevant to mitochondrial biogenesis, such as PPARc coactivatorsPGC-1a and PGC-1b) and mitochondrial transcription factor A (Tfam) in SMS1-KO WAT relative to wild-type tissues (Fig. 4C, Table S1). To confirm these findings, we assessed expression of components of the mitochondrial electron transport complex and ATP synthase (Fig. 4D, Table S1). Some of the former, such as as9 (NDUFA9) and.E significant increment of caspase-3 (a downstream apoptotic factor) activity (Fig. 3E). We also observed increased expression of macrophage markers (F4/80 and CD68) in SMS1KO WAT, probably suggesting enhanced invasion of macrophage (Fig. 3F).Hyperlipidemia and Lipodystrophy in SMS1-KO Mice Are Rescued by Anti-oxidant TreatmentOur results support the idea that WAT cells of SMS1-KO mice are functionally damaged by increased oxidative stress. Indeed, triglyceride levels were greatly increased in blood plasma of SMS1KO compared to wild-type mice, indicative of hyperlipidemia (Fig. 6A, also see Fig. 2A). In particular, triglyceride levels of chylomicron (CM) and very-low-density lipoprotein (VLDL), which normally function in muscle or are stored in adipocytes [43], were greatly increased in blood plasma, suggesting that lipid storage function of adipose cells is compromised in SMS1-KO mice (Fig. 6A). To further examine whether oxidative stress mediates these phenotypes, we supplied the anti-oxidant N-acetyl cysteine (NAC) in the drinking water of mutant mice and analyzed potential changes in blood plasma triglyceride levels (Fig. 6A). NAC-treated SMS1-KO mice showed robust decreases in blood triglyceride levels, almost to levels comparable to wild-type mice. Finally, we subjected mice to fasting for 30 hours and assessed triglyceride levels in blood plasma (Fig. 6B). Those levels increased in fasting wild-type mice and in SMS1-KO mice supplied normal drinking water. However, CM levels in fasting SMS1-KO mice were significantly decreased compared to wild-type mice. By contrast, SMS1-KO mice supplied with NAC-containing drinking water exhibited partially recovered CM levels. The volume of SMS1-KO epiWAT was also partially rescued by NAC-treatment (Fig. 6C). Altogether, these results indicate that increased oxidative stress underlies a failure of lipid storage function in adipocytes of SMS1-KO mice.Oxidative Stress Response, the Mitochondrial Stress Response and Mitochondrial Biogenesis Are Enhanced in SMS1-KO WATTo confirm whether oxidative stress occurs in SMS1-KO WAT, we examined expression of mRNAs encoding ROS detoxification enzymes. In SMS1-KO WAT, expression of some isoforms of peroxiredoxin (Prdx), which reduces hydrogen peroxide and alkyl hydroperoxides, was increased (Fig. 4A, Table S1). Expression of catalase, which catalyzes decomposition of hydrogen peroxide, also increased. Other ROS detoxification enzymes, such as glutathione peroxidase (Gpx), 15755315 superoxide dismutase (SOD) and glutaredoxin (Glrx), also showed increased expression. These data indicate that the oxidative stress response is enhanced in SMS1KO WAT. Next we examined expression of mRNAs encoding mitochondrial stress response-related factors. Most of these, including mitochondrial chaperones (mtHsp70, mtHsp40) and mitochondrial proteases (LONP1, ClpP, YME1L1, AFG3L2), were upregulated in SMS1-KO WAT (Fig. 4B, Table S1). On the other hand, mRNA expression of calnexin, an ER chaperone, or HSP70-1, a cytosolic chaperone, was not affected. We also observed increases in mRNAs encoding factors relevant to mitochondrial biogenesis, such as PPARc coactivatorsPGC-1a and PGC-1b) 
and mitochondrial transcription factor A (Tfam) in SMS1-KO WAT relative to wild-type tissues (Fig. 4C, Table S1). To confirm these findings, we assessed expression of components of the mitochondrial electron transport complex and ATP synthase (Fig. 4D, Table S1). Some of the former, such as as9 (NDUFA9) and.
