Figure 9. Rab18/NRZ/SNARE complex establishes ER–LD contact. (A) Presence of ER-associated SNAREs in LD fraction is dependent on NAG. Subcellular fractions were isolated from control or NAG KO 3T3-L1 preadipocytes. (B and C) Use1 (green) was colocalized with Rab18 (red) to LDs (blue) in the control but not in NAG KO or ZW10 KO 3T3-L1 preadipocytes. Bars: 5 µm; (insets) 2 µm. (C) Quantitative analysis. Cells with Rab18 were selected for calculation and LDs with GFP-Use1 localization were counted as positive. Three independent experiments were performed. Mean ± SD; n = 3; ***, P < 0.001; NS, no significance by one-way ANOVA with Tukey post hoc tests. (D) Colocalization of CB5 (green, an ER-associated protein), Rab18 (red), and BNIP1 (pink) on LDs in wild-type but not in NAG KO cells. Bars: 10 µm; (insets) 2 µm. (E) Quantification of percentage of cells containing CB5 positive LDs in D. Rab18-positive cells were selected for calculation. Three independent experiments were performed. Mean ± SD; n = 3; ***, P < 0.001 by two-tailed t test. (F) Colocalization of GFP-Use1 (green), RFP-KEDL (red, an ER-specific marker) and HA-Rab18 (pink) on LDs. 3D view of indicated channels are shown in xy, xz, or yz direction. Bars: 10 µm; (insets) 2 µm. All experiments were performed at least twice.
Figure 9.

Rab18/NRZ/SNARE complex establishes ER–LD contact. (A) Presence of ER-associated SNAREs in LD fraction is dependent on NAG. Subcellular fractions were isolated from control or NAG KO 3T3-L1 preadipocytes. (B and C) Use1 (green) was colocalized with Rab18 (red) to LDs (blue) in the control but not in NAG KO or ZW10 KO 3T3-L1 preadipocytes. Bars: 5 µm; (insets) 2 µm. (C) Quantitative analysis. Cells with Rab18 were selected for calculation and LDs with GFP-Use1 localization were counted as positive. Three independent experiments were performed. Mean ± SD; n = 3; ***, P < 0.001; NS, no significance by one-way ANOVA with Tukey post hoc tests. (D) Colocalization of CB5 (green, an ER-associated protein), Rab18 (red), and BNIP1 (pink) on LDs in wild-type but not in NAG KO cells. Bars: 10 µm; (insets) 2 µm. (E) Quantification of percentage of cells containing CB5 positive LDs in D. Rab18-positive cells were selected for calculation. Three independent experiments were performed. Mean ± SD; n = 3; ***, P < 0.001 by two-tailed t test. (F) Colocalization of GFP-Use1 (green), RFP-KEDL (red, an ER-specific marker) and HA-Rab18 (pink) on LDs. 3D view of indicated channels are shown in xy, xz, or yz direction. Bars: 10 µm; (insets) 2 µm. All experiments were performed at least twice.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal