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[Preprint]. 2023 Oct 14:2023.10.13.562302. [Version 1] doi: 10.1101/2023.10.13.562302

Figure 1. Constitutive endogenous expression of PNPLA3-I148M increases the LD content of hepatoma cells.

Figure 1.

(A) PNPLA3-I148M was introduced into Hep3B cells using CRISPR. Parental Hep3B cells (WT and I148M) were further modified to introduce a C-terminal HiBiT tag at the endogenous locus. (B) PNPLA3 expression in the parental cells (left) was analyzed by digital PCR (p = 0.0972, determined by Student’s t-test, n = 3). Protein expression (right) was analyzed by HiBiT detection in the tagged cell lines treated with a negative control siRNA or PNPLA3-specific siRNA. (C,D) Mean lipid droplet content (upper) and mean lipid droplet area (lower) were quantified from live-cell label-free imaging using the Nanolive 3D Cell Explorer microscope. Cells were treated with vehicle (C) or with 200 μM oleic acid (D). Images were taken every 3 min for 24 h, starting 1 h after addition of oleic acid. Quantification was done using Eve software.