Fig 1: DOP1A targets Agpat2 to regulate PL synthesis(A) Western blot of the cytoplasmic fraction enriched in ER components and the nuclear fraction containing NM components in Neuro2a cells.(B) PL profiles of the nuclear fraction. The nuclei of control and Dop1a knockdown cells cultured in the presence of Torin1 were subjected to LC-MS. Values are shown as Z scores calculated for each lipid species across all samples.(C) Visualization of nuclear lipid droplets (nLDs) in the Dop1a-depleted cells treated with Torin1. The nLDs were stained with Bodipy 493/503 (BD493). Scale bars, 10 μm.(D) Frequency of cells exhibiting nLDs (n = 267 and 55 cells).(E) The position of nLD within the nucleus observed by the z axis stacking.(F) The frequency of nLDs is showing continuity with the NM (n = 101 and 99 cells).(G) Super-resolution microscopy of Agpat2 and Nup153. The linescan was performed along the dashed line indicated in the inset image. Scale bars, 2 μm.(H) Coloc2 analysis of Agpat2 and Nup153 fluorescent signals in the cells treated with Torin1 or DMSO. PCC, Pearson’s correlation coefficient; n.s., not significant.(I) The PL global analysis performed in Neuro2a cells after depletion of both Dop1a and Agpat2, followed by Torin1 treatment. Values are shown as Z scores calculated for each lipid species across all samples.(J) Schematic for the coregulatory module of Dop1a and Agpat2 over the PLs. Data are represented as means ± SEM. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001, unpaired two-tailed Student’s t test.