Figure 1. Defective LD growth and... | Rockefeller University Press

$Figure 1. Defective LD growth and maturation in Rab18-deficient cells. (A) Outline of strategy to screen for LD-associated Rabs involved in LD growth. (B) Knocking down Rab18 alters LD morphology in 3T3-L1 preadipocytes. Bars: 10 µm; (insets) 2 µm. (C) Representative images of LDs (green) in negative control (NC) or Rab18-deficient (Rab18 KO) 3T3-L1 preadipocytes. Red represents Cherry expression. Bars: 10 µm; (insets) 2 µm. (D) Histogram showing the mean number of LDs in each diameter in C. Data represent mean ± SD (n = 24 cells for NC; n = 25 cells for Rab18 KO; n = 29 cells for Rab18 KO+Rab18; ***, P < 0.001; NS, no significance by Kruskal-Wallis test). (E) Schematic diagram showing the process of LD biogenesis and growth. (F–H) Quantification of the number of mature LDs under fluorescent microscope (F), the percentage of LDs in each diameter (G), and diameter of the largest LD in each cell (H) in C. Mean ± SD for F, mean value for G, mean ± SEM for H, n = 24 cells for NC; n = 25 cells for Rab18 KO; n = 29 cells for Rab18 KO+Rab18; ***, P < 0.001; NS, no significance by Kruskal-Wallis test. (I) Reduced total TAG levels in Rab18 KO 3T3-L1 preadipocytes. TAG/protein level of control cells was normalized to 1. Three independent experiments were performed. Mean ± SD; n = 3; **, P < 0.01 by two-tailed t test. (J) Relative TAG level in various subcellular fractions. TAG level in PNS in control cells was normalized to 1. Three independent experiments were performed. Mean ± SD; n = 3; *, P < 0.05; **, P < 0.01; NS, no significance by two-tailed t test. (K and L) Increased BIP expression and mRNA levels of spliced XBP-1 in Rab18 KO 3T3-L1 preadipocytes after OA (400 µM) treatment. Cells treated with 1 µM TG for 6 h were used as a positive control. Three independent experiments were performed. Mean ± SD; n = 3; *, P < 0.05; **, P < 0.01 by two-tailed t test. All experiments were performed at least twice. WT, wild type.$

Figure 1.

Defective LD growth and maturation in Rab18-deficient cells. (A) Outline of strategy to screen for LD-associated Rabs involved in LD growth. (B) Knocking down Rab18 alters LD morphology in 3T3-L1 preadipocytes. Bars: 10 µm; (insets) 2 µm. (C) Representative images of LDs (green) in negative control (NC) or Rab18-deficient (Rab18 KO) 3T3-L1 preadipocytes. Red represents Cherry expression. Bars: 10 µm; (insets) 2 µm. (D) Histogram showing the mean number of LDs in each diameter in C. Data represent mean ± SD (n = 24 cells for NC; n = 25 cells for Rab18 KO; n = 29 cells for Rab18 KO+Rab18; ***, P < 0.001; NS, no significance by Kruskal-Wallis test). (E) Schematic diagram showing the process of LD biogenesis and growth. (F–H) Quantification of the number of mature LDs under fluorescent microscope (F), the percentage of LDs in each diameter (G), and diameter of the largest LD in each cell (H) in C. Mean ± SD for F, mean value for G, mean ± SEM for H, n = 24 cells for NC; n = 25 cells for Rab18 KO; n = 29 cells for Rab18 KO+Rab18; ***, P < 0.001; NS, no significance by Kruskal-Wallis test. (I) Reduced total TAG levels in Rab18 KO 3T3-L1 preadipocytes. TAG/protein level of control cells was normalized to 1. Three independent experiments were performed. Mean ± SD; n = 3; **, P < 0.01 by two-tailed t test. (J) Relative TAG level in various subcellular fractions. TAG level in PNS in control cells was normalized to 1. Three independent experiments were performed. Mean ± SD; n = 3; *, P < 0.05; **, P < 0.01; NS, no significance by two-tailed t test. (K and L) Increased BIP expression and mRNA levels of spliced XBP-1 in Rab18 KO 3T3-L1 preadipocytes after OA (400 µM) treatment. Cells treated with 1 µM TG for 6 h were used as a positive control. Three independent experiments were performed. Mean ± SD; n = 3; *, P < 0.05; **, P < 0.01 by two-tailed t test. All experiments were performed at least twice. WT, wild type.

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