Fig. 6

Mechanistic target of rapamycin complex 1 (mTORC1) signaling remains unrepressed in the fasted pex5^−/− liver. a Western blotting shows the expression levels of 4EBP and its phosphorylated form from whole lysates of WT and pex5^−/− zebrafish at 6 dpf. The expression of β-actin was used as the loading control. Graph shows quantified expression levels of p4EBP relative to β-actin levels and presented as the average of three repeats with error bars indicating standard deviation. Statistical significance was determined using the Student’s t test in Microsoft Excel; * indicates p value < 0.05. b Immunofluorescence was used to detect the differential levels of S6 and its phosphorylated form in the liver cells of WT and pex5^−/− at 6 dpf. DAPI detects nuclei in each liver section. Scale bar equals to 5 μm. Graph shows relative fluorescence intensity presented as the average with error bars indicating standard deviation. Statistical significance was determined using the Student’s t test in Microsoft Excel; ** indicates p values < 0.01. c Rapamycin (rapa, 250 nM) was used to treat WT and pex5^−/− from 3.5 to 8 dpf and representative images of the resulting larvae were laterally presented with anterior to the left. The average percentage representing a near-complete rescue from two independent experiments is indicated. Scale bar = 1 mm. d Liver sections from the larvae treated with rapamycin from 3.5 to 7 dpf were stained either ORO or PAS. Representative images are shown, and the percentage of rescue in the levels of liver lipids and glycogen are indicated in the panels of the fasted pex5^−/− zebrafish treated with rapamycin. Scale bar = 100 μm