Figure 2. Rab18 deficiency causes abnormal LD growth in adipocytes and Leydig cells. (A and B) Knocking down Rab18 alters LD (green) sizes in adipocytes. Bars: 10 µm; (insets) 5 µm. (B) Size distribution of the largest LD. LDs from 88 siRab18-transfected cells and 116 siNC-transfected cells were analyzed (pooled from three experiments). LD size distribution was fitted with Gaussian function. (C and D) Overexpressing Rab18 decreases LD sizes in adipocytes. Green, GFP; red, LDs. Bars: 10 µm; (insets) 5 µm. (D) The size distribution of the largest LD. LDs from 121 Rab18-transfected cells and 100 vector-transfected cells were analyzed (pooled from three experiments). LD size distribution was fitted with Gaussian function. (E) Relative cellular TAG levels in adipocytes. TAG/protein level of siNC was normalized to 1. Three independent experiments were performed. Mean ± SD; n = 3; ***, P < 0.001 by two-tailed t test. (F) Reduced basal and stimulated lipolysis in Rab18 knockdown adipocytes. The amount of free glycerol released from siNC cells without the treatment of isoproterenol (iso) and 3-isobutyl-1-methylxanthine (IBMX) was normalized to 1. Three independent experiments were performed. Mean ± SD; n = 3; **, P < 0.01; ***, P < 0.001 by one-way ANOVA with Tukey post hoc tests. (G and H) Rab18 deficiency in TM-3 Leydig cell alters LD morphology. Green, LDs. Bars: 5 µm; (insets) 2 µm. (H) Histogram showing the mean number of LDs in G. Mean ± SD; n = 10; ***, P < 0.001; NS, no significance by two-tailed t test. All experiments were performed at least twice.
Figure 2.

Rab18 deficiency causes abnormal LD growth in adipocytes and Leydig cells. (A and B) Knocking down Rab18 alters LD (green) sizes in adipocytes. Bars: 10 µm; (insets) 5 µm. (B) Size distribution of the largest LD. LDs from 88 siRab18-transfected cells and 116 siNC-transfected cells were analyzed (pooled from three experiments). LD size distribution was fitted with Gaussian function. (C and D) Overexpressing Rab18 decreases LD sizes in adipocytes. Green, GFP; red, LDs. Bars: 10 µm; (insets) 5 µm. (D) The size distribution of the largest LD. LDs from 121 Rab18-transfected cells and 100 vector-transfected cells were analyzed (pooled from three experiments). LD size distribution was fitted with Gaussian function. (E) Relative cellular TAG levels in adipocytes. TAG/protein level of siNC was normalized to 1. Three independent experiments were performed. Mean ± SD; n = 3; ***, P < 0.001 by two-tailed t test. (F) Reduced basal and stimulated lipolysis in Rab18 knockdown adipocytes. The amount of free glycerol released from siNC cells without the treatment of isoproterenol (iso) and 3-isobutyl-1-methylxanthine (IBMX) was normalized to 1. Three independent experiments were performed. Mean ± SD; n = 3; **, P < 0.01; ***, P < 0.001 by one-way ANOVA with Tukey post hoc tests. (G and H) Rab18 deficiency in TM-3 Leydig cell alters LD morphology. Green, LDs. Bars: 5 µm; (insets) 2 µm. (H) Histogram showing the mean number of LDs in G. Mean ± SD; n = 10; ***, P < 0.001; NS, no significance by two-tailed t test. All experiments were performed at least twice.

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