Journal: Nature

Article Title: CLCC1 promotes hepatic neutral lipid flux and nuclear pore complex assembly

doi: 10.1038/s41586-025-10064-4

Figure Lengend Snippet: A) Histogram indicating the distribution of control (null) sgRNAs. Shown below is the relative enrichment of CLCC1 targeting sgRNAs (red lines) under untreated, triacsin C, oleate, and serum starve conditions. The gray line indicates the mean of the control sgRNA distribution. B) Representative flow cytometry histograms of control and CLCC1 KO Huh7 cells following treatment with 1 µg/mL triacsin C for 24 h, 100 µM oleate for 24 h, serum starve for 48 h, or basal levels. C) Representative confocal images of lipid droplets using BODIPY 493/503 in control (expressing safe targeting sgRNA) and CLCC1 KO cells under 100 µM oleate for 24 h or serum starve for 48 h (also see Fig. for imaging of untreated and triacsin C treated cells). D) Quantification of the number of LDs from images shown in Fig. and Extended Data Fig. 2c. Data represent mean ± SD of > 100 cells across three biological replicates. E) Quantification of the area of LDs from images shown in Fig. and Extended Data Fig. 2c. Data represent mean ± SD of > 100 cells across three biological replicates. F) Representative transmission EM images of negative stained control and CLCC1 KO Huh7 cells. G) Top: TLC resolving of TAG, CE, and polar lipids in control and CLCC1 KO Huh7 cells. Bottom: Quantified TAG (left graph) and CE (right graph) bands normalized to phospholipids (PL). Data represent mean ± SD of three biological replicates. H) Immunoblot of three Clcc1 fl/fl mice injected with either AAV-GFP (control) or AAV-Cre (Clcc1 HepKO ). Samples were analyzed four weeks post-injection. I) Representative transmission EM images of negative stained control and Clcc1 HepKO mice. J,K) Plasma fractionation by FPLC from male control and Clcc1 HepKO mice measuring (J) cholesterol in all fractions and (K) TAGs in all fractions. (see Fig. for female mouse data) L) Immunoblot of apoB48 and apoB100 in indicated fractions for VLDL-associated apoB and LDL-associated apoB. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; NS, not significant ( P ≥ 0.05) by one-way ANOVA with Dunnett’s multiple comparisons test.

Article Snippet: Size-exclusion chromatography was performed using an AKTA FPLC (Amersham Pharmacia Biotech).

Techniques: Control, Flow Cytometry, Expressing, Imaging, Transmission Assay, Staining, Western Blot, Injection, Clinical Proteomics, Fractionation

Journal: Nature

Article Title: CLCC1 promotes hepatic neutral lipid flux and nuclear pore complex assembly

doi: 10.1038/s41586-025-10064-4

Figure Lengend Snippet: a , Representative images of livers from control and Clcc1 -HepKO mice. b , Change in liver mass normalized to body mass for control and Clcc1 -HepKO mice. n > 8. c , Body mass of the indicated control and Clcc1 -HepKO mice. n > 8. d , e , Quantification of TAG ( d ) and cholesteryl ester (CE) ( e ) normalized to phospholipid (PL) content using thin layer chromatography (TLC). Data represent mean ± s.d. of six mice. f , Representative haematoxylin and eosin (H&E), oil red O, Masson’s trichrome and picrosirius red-stained liver sections from control and Clcc1 -HepKO mice. Scale bar, 100 μm. g , h , Plasma fractionation by fast protein liquid chromatography (FPLC) from female control and Clcc1 -HepKO mice measuring cholesterol ( g ) and TAG ( h ) in all fractions. LDL, low density lipoprotein. i , Western blot analysis of apoB and albumin in plasma from four female control mice and four Clcc1 -HepKO mice. j , Quantification of TAG in plasma using TAG-Glo Assay (Promega, J3160). Data represent mean ± s.d. of six mice. k , Quantification of AST in serum of wild-type and Clcc1 -HepKO mice. Data represent mean ± s.d. of more than four mice. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; NS, not significant ( P ≥ 0.05) by one-way ANOVA with Dunnett’s multiple comparisons test.

Article Snippet: Size-exclusion chromatography was performed using an AKTA FPLC (Amersham Pharmacia Biotech).

Techniques: Control, Thin Layer Chromatography, Staining, Clinical Proteomics, Fractionation, Fast Protein Liquid Chromatography, Western Blot, Glo Assay