Extended Data Fig. 6: Construction of SREBP2 (F167A) knock-in mice.
a, Alignment of the amino acid sequence of nSREBP2s from different species. Sequence alignment shows that F178 in human SREBP2 is a conserved residue back to killifish. b, Strategy to generate F167A knock-in mice by CRISPR/Cas9 system. c, DNA sequencing results of F167 residue of WT and F167A mice. d, Western blotting showing that F167A mutation did not affect the SREBP2 cleavage process in WT control and F167A knock-in mice primary hepatocytes (n = 3 mice per group). e, Reproductive strategy to generate F167A knock-in homozygous mice. f, Immunofluorescence analysis of SREBP2 WT and F167A mutation mice primary hepatocytes. After cholesterol depletion for 16 h, primary hepatocytes were stained and then imaged with Nikon CSU-W1 confocal microscope. Scale bars for the large images and zoomed images are 25 μm and 5 μm. Imaging data represents three independent experiments. g, Quantification of the percentage of cells that displayed nuclear puncta is shown in (f). Data were shown as mean ± s.d. 100 cells in each group were quantified; n = 3 biologically independent samples. Statistical significance was calculated with unpaired two-tailed Student’s t-test.
