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. 2021 Oct 21;134(20):jcs258894. doi: 10.1242/jcs.258894

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© 2021. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 8. — Interaction of sortilin and FABP7 in PPARγ-dependent gene expression. (A) CHO cells stably expressing sortilin (CHO-S) were transfected with reporter gene constructs encoding a PPAR-responsive firefly luciferase gene (Luc) and a constitutively expressed Renilla luciferase gene (Ren). Where indicated, transfectants were also treated with rosiglitazone (+RGS) for 24 h. At 48 h after transfection, the activities of firefly and Renilla luciferases were determined in cell lysates using a luminometer (n=3–8 replicates per cell line). Values are given as ratio of firefly to Renilla luciferase activity (mean±s.e.m., condition without RGS as 100%). ***P<0.001; ****P<0.0001 (unpaired, two-tailed Student's t-test). (B) Replicate layers (n=4) of CHO cells stably expressing sortilin (CHO-S), FABP7 (CHO-F), or both proteins (CHO-S/F) were transfected with constructs encoding a PPAR-responsive firefly luciferase and a constitutively expressed Renilla luciferase and analyzed for luciferase activity as described above. Values are given as ratio of firefly to Renilla luciferase (mean±s.e.m., CHO-S set to 100%). **P<0.01; ***P<0.001 (unpaired, two-tailed Student's t-test). (C) CHO-S/F were transfected with reporter gene constructs encoding a PPAR-responsive firefly luciferase reporter gene and a constitutively expressed Renilla luciferase. Then, transfectants were treated with conditioned medium containing 5 µg/ml of human apoE3 or apoE4 for 24 h. At 48 h after transfection, the activities of firefly and Renilla luciferases were determined in cell lysates using a luminometer (n=9 replicates from 3 independent experiments per condition. Values are given as ratio of firefly to renilla luciferase (mean±s.e.m., +apoE3 set to 100%). *P<0.05 (unpaired, two-tailed Student's t-test). (D,E) Proposed model of the interaction of sortilin with FABP7 and apoE3 (D) or apoE4 (E) in cellular lipid metabolism (see Discussion section for details).