Fig. 5: PNPLA6 knockdown in primary human RPE cells disturbs choline turnover.
From: PNPLA6 regulates retinal homeostasis by choline through phospholipid turnover
a qPCR analysis of PNPLA6 in PNPLA6-knockdown (si PNPLA6) and control (scramble) primary human RPE cells, with GAPDH as an internal control (n = 3 biological replicates). b, c Lipidomics of PC (n = 4 biological replicates) (b) and LPC (n = 3 biological replicates) (c) in PNPLA6-knockdown and control cells. d Changes in choline level in PNPLA6-knockdown cells relative to control cells, with the levels in control cells as 1 (n = 5 biological replicates). e Proliferation of PNPLA6-knockdown cells relative to control cells, with the levels in control cells as 1 (n = 3 biological replicates). f TERR analysis of PNPLA6-knockdown and control cells (n = 3 biological replicates). g Phagocytosis of FITC-labeled (red) porcine POS in PNPLA6-knockdown and control cells. Scale bar, 10 μm. h Quantitative values of phagocytosis, with control cells set as 1 (n = 4 biological replicates). i JC-1 staining of PNPLA6-knockdown and control cells. Scale bar, 10 μm. j Red/green ratio of JC-1 staining (n = 4 biological replicates). k, l TEM images of PNPLA6-knockdown and control cells. Representative images (left) with quantification of mitochondria number (n = 5 biological replicates) (right) (k) and microvilli (n = 5 biological replicates) (l) are shown. Arrows, mitochondria. Scale bar, 1 μm. (m) Effects of choline and CHKα inhibitor (MN58b) on the proliferation of PNPLA6-knockdown cells (n = 8 biological replicates). Data are presented as mean ± SEM and representative of at least two independent experiments. P-values were derived from one-way ANOVA with Tukey’s multiple comparison test (m) and two-tailed unpaired-t test (a–f, h, j, k, and l). Source data are provided as a Source Data file.
