Fig 2: RPE and retinal functionality in RPE-Pnpla2-cKO mice. (A) ERG amplitude graphs of scotopic a- and b-waves and photopic b-waves, as a function of light intensity (x-axis) of 3- and 12-month-old cKO mice (open circles) and littermate controls (Pnpla2f/f, closed circles) (n = 3 per genotype). (B) Bar graph showing the amplitude (mean, SD) of the c-wave, fast oscillation (FO), light peak (LP), and off-response (OFF) measured by DC-ERG of 11-week-old cKO mice (n = 4, open bars) and Pnpla2f/f and Pnpla2f/+ control mice (n = 5, closed bars).

Fig 3: PEDF decreases H/R-induced apoptosis via PEDF-R in cultured H9c2 cells. H9c2 cells were maintained in normoxic or H/R conditions for 8/2 h with or without PEDF (10 nM). RNA interference assays were used to silence PEDF-R. (A) Samples were collected for western blotting to analyze the expression of cleaved casp3 protein, with (B) quantification performed using ImageJ software (n=4). (C) Effect of PEDF on H9c2 cells apoptosis, with (D) quantification. TUNEL (red) staining was performed for each group. Nuclei were stained with DAPI (blue). Cells that were TUNEL and DAPI-positive were apoptotic (indicated by the arrows), while DAPI positive were control cells (scale bar=50 µm; n=4). Data are expressed as the mean ± standard error of the mean. *P<0.05, with comparisons indicated by lines. PEDF, pigment epithelium-derived factor; H/R, hypoxia/reoxygenation; PEDF-R, pigment epithelium-derived factor receptor; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling; si, small interfering.

Fig 4: LBP significantly reduces hepatic triglyceride accumulation in cells chronically exposed to PA.(A,B) Cells were treated for 24 hours with different concentrations of LBP in the absence or presence of PA. ATGL and FAS protein expressions were examined by immunoblotting. All data shown are representative of three independent experiments. (C) Cells were treated with LBP and exposed to PA followed by Oil red O staining. (Scale bar: 7.5 µm). (D,E) Cells transfected by siRNA were treated with LBP in absence or presence of PA. All data shown are representative of three independent experiments. (F) After transfection by siRNA, cells were treated for 24 hours with LBP in the absence or presence of PA. Next, the triglyceride levels in each group were measured. Data are expressed as the mean ± SEM (n = 5). Note: n.s, non-significant.

Fig 5: Lipid accumulation in the RPE of Pnpla2-cKO mice. Electron microscopy micrographs showing the RPE structure of 3-month-old (A) and 13-month-old (B) cKO mice and control animals. Scale bar: 2 µm. The representative images were selected among examinations of micrographs from eight eyes of cKO mice (PNPLA2f/f/Cre+), from seven eyes of PNPLA2f/f control mice at 1.75 to 3.75 months old, and from three eyes of cKO mice and three eyes of control mice at 12.5 to 13 months old. LD, lipid droplets; BI, basal infoldings.