ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Expression patterns of zebrafish soat1 during embryogenesis.

Whole-mount in situ hybridization shows that soat1 can be detected in all blastomeres at 1 cell stage (A), 3 hpf (B), and 6 hpf (C). The expression of soat1 was prominent at yolk sac at 12 hpf (D) and 24 hpf (E). By 48 hpf (F, G), soat1 was detected in the brain, retina, pectoral fin, hatching gland and pericardium. By 72 hpf (H, I), soat1 was also observed prominently in the liver primordia and intestine.

Expression patterns of zebrafish soat2 during embryogenesis.

Whole-mount in situ hybridization shows that soat2 can be detected at yolk sac at 12 hpf (A) and 24 hpf (B). By 48 hpf (C, D), soat2 was detected in the brain, retina, yolk sac, hatching gland, pericardium and intersomitic regions. By 72 hpf (E, F), soat2 was also observed in the liver primordial and intestine.

Increased intracellular accumulation of neutral lipid and CEs as a result of esterification of cholesterol to fatty acyl-CoA.

(A) Oil Red O (ORO) staining of eGFP stable expressing (Ctrl), zebrafish saot1 stable expressing (Soat1), and zebrafish soat2 stable expressing (Soat2) HEK293 cells after incubation with low (15 μM oleic acids and 1 μg/mL cholesterol) or high (150 μM oleic acids and 10 μg/mL cholesterol) level of substrate at room temperature for 6 hrs. The expression of zebrafish Soat2 significantly increased the intracellular accumulation of neutral lipid indicated its enzymatic activity of cholesterol esterification, and this accumulation of neutral lipid is significantly increased with the increased levels of substrates. Note the significantly increased neutral lipid in WT-high compared to WT-low reflected the activity of endogenous human SOAT1 expressed by HEK293 cells. The letters above each column indicate the statistical groups, and the data sharing the same letters indicates no significant difference. (B) The cell lysate of HEK293 cells with zebrafish soat2 overexpression contained significantly higher CE content than either control group or soat1 overexpression group. Consistent with the ORO staining, the CE content in soat1 group was comparable to the control group. (C) RT-PCR confirmed the expression of both zebrafish soat1 and eGFP in Soat1-overexpressing HEK293 cells, zebrafish soat2 and eGFP in Soat2-overexpressing HEK293 cells and only eGFP in control group in which the HEK293 cells were transfect with empty vectors. (D) The HEK293 cells overexpressing zebrafish Soat2 (Soat2-DsRed) contained more and larger intracellular lipid droplets containing NBD-cholesterol as compared to the HEK293 cells overexpressing DsRed only (DsRed). (E) Avasimibe (AVA) significantly reduced the intracellular accumulation of neutral lipid, but the cells with zebrafish soat2 overexpression still showed a significantly more ORO staining than other AVA-treated cells. (F) Pyripyropene A (PPPA) could significantly reduce the intracellular accumulation of neutral lipid by inhibiting the enzymatic activity of both endogenous human SOAT1 and zebrafish Soat2 when compared to the vehicle control (DMSO). The letters above each column indicate the statistical groups, and the data sharing the same letters indicates no significant difference.

Delayed yolk consumption by yolk injection of PPPA.

After the injection of PPPA, AVA or DMSO into the yolk at 3 hpf, embryos with grossly normal appearance at 24 hpf were subjected to yolk area measurement to estimate the yolk consumption. Yolk injection of PPPA resulted in significant larger yolk size at 72 and 84 hpf when compared to the DMSO group suggesting a delayed consumption of yolk content. *: P < 0.05, PPPA vs. DMSO; ***: P < 0.001, PPPA vs. DMSO; #: P < 0.05, PPPA vs. AVA

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Zebrafish Soat2 catalyzes the esterification of cholesterol.

After 2- and 5-hour incubations with 150 μM oleic acids, 10 μg/mL cholesterol and 10 μg/mL NDB-cholesterol, the intracellular accumulation of CEs was observed in the HEK293 cells transiently overexpressing DsRed, zebrafish Soat1-DsRed and zebrafish Soat2-DsRed. The progress of CE accumulation could be seen in cells with zebrafish Soat2 overexpression.