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. Author manuscript; available in PMC: 2020 Oct 22.
Published in final edited form as: Biochim Biophys Acta Mol Cell Biol Lipids. 2016 Dec 23;1862(3):358–368. doi: 10.1016/j.bbalip.2016.12.009

Fig. 5.

Fig. 5

Transient silencing of Pex16 in mature 3T3-L1 adipocytes (A)–(H) Transient silencing of Pex16 in mature 3T3-L1 adipocytes (day 5 of differentiation) by electroporation (EP) of 200 nM control siRNA (si-ctrl) or siRNA directed against Pex16 (si-Pex16). (A) RT-PCR analysis of Pex16 expression 48 h, 60 h and 72 h after EP of 3T3-L1 adipocytes with siRNA. (B)–(H) Data analyzed from si-ctrl and si-Pex16 3T3-L1 adipocytes at day 7 (=48 h after EP). (B) RT-PCR analysis of peroxisomal genes (Pex16, Pex11b, Pex14, Catalase, Gnpat), adipogenesis marker genes (Pparγ, aP2) and fatty acid metabolism genes (Cd36, Fas). (C) Protein expression and (D) densitometric analysis of PEX16, PEX14 and fatty acid synthase (FAS). (E) Relative number of peroxisomes counted from ELMI pictures. (F) Catalase activity. (G) 14C–oleic acid (OA) uptake into si-ctrl and si-Pex16 3T3-L1 cells after 30 min incubation. (H) Oxygen consumption rate in the presence of 10 μM hexacosanoic acid (C26:0). Cells were incubated with assay medium containing 10 μM C26:0 45 min prior to and during the measurement with Seahorse XF96 extracellular flux analyzer (n = 4). (A),(B),(D)–(G) Data are presented as mean ± SD (n = 3). (H) Data are presented as mean ± SEM. Statistical significance was calculated using Student’s t-test. *p < 0.05, ** p < 0.01, ***p < 0.001 versus control.