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. 2019 Apr 9;69(6):2427–2441. doi: 10.1002/hep.30583

Figure 2.

Figure 2

Cellular immunolocalization of PNPLA3(WT and 148M) in QBI‐293A cells. (A) QBI‐293A cells were transfected with the plasmids encoding human PNPLA3(WT or 148M) and then cultured for 24 hours in the presence or absence of 200 µM OA. Immunofluorescence microscopy was performed as described in Fig. 1. LDs were stained using MDH (cyan, left). White dash lines outline the transfected cells. Protein expression was analyzed by immunoblotting using lactate dehydrogenase as a loading control (right). (B) The experiment was repeated using PNPLA3 with a mC epitope tag at the N‐ter or C‐ter in the presence of 200 µM OA. (C) The experiment described in (A) was repeated in the presence of OA with the inclusion of cells expressing mC alone. Sections were analyzed by ImageJ to calculate total LD areas. Diameters of the 10 largest LDs from each cell were measured in 18 or more cells. Tukey's multiple comparisons test; **P < 0.01, ***P < 0.001, ****P < 0.0001. Scale bar = 10 µm. All experiments were repeated twice and yielded similar results. Abbreviation: LDH, lactate dehydrogenase.