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. Author manuscript; available in PMC: 2019 May 1.
Published in final edited form as: Cell Metab. 2018 May 1;27(5):1138–1155.e6. doi: 10.1016/j.cmet.2018.03.015

Figure 4. Functional validation of Lcn5 as a regulator of skeletal muscle metabolism in vitro.

Figure 4

A, qPCR analysis of Lcn5 expression across indicated tissues in C57BL6J mice. B, Suggestive pathway enrichment derived from skeletal muscle genes ranked by correlation with adipose Lcn5. C, Adipose Lcn5 expression correlation with clinical traits within the HMDP. D, C2C12 myotubes were treated overnight with media containing either PBS (veh) or recombinant LCN5 protein (1μg/mL) then subjected to qPCR analysis. E–H, The same treatment as in (D) was carried out for 30hrs, then cells were immunoblotted for respiratory complex abundance (E) or subjected to Seahorse bioanalyzer (F) to assess oxygen consumption in both basal (G) and maximal respiratory (H) conditions. For Seahorse experiments, oxygen consumption was normalized to total protein content. I–K, 3T3l1 preadipocytes infected with either AAV-GFP or AAV-LCN5 were differentiated and placed in cocultures with C2C12 myotubes (I), AML12 hepatocytes (J) or 3T3L1 adipocytes (J) for 24 hrs and qPCR-probed for Sdhc or Atp5f1. n=4–8 per group. All data presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001