FIGURE 7.

mRNA expression changes of Mfsd14A and Mfsd14B in cells and tissue samples. Mouse cortex primary cultures from e14.5-15 were subjected to amino acid starvation and the mRNA levels monitored at 3, 7 and 12 h. Normalization was performed to three housekeeping genes (Gapdh, H3a and Actb). The relative mRNA expression (±SD) was plotted, with the highest relative expression set to 100%. Expression differences were calculated using t-test and Bonferroni corrected significance levels (^∗p < 0.0493, ^∗∗p < 0.00998, ^∗∗∗p < 0.001). (A) Mfsd14a expression in amino acid starved cells (AA starved) as compared to control cells and (B) Mfsd14b expression compared to control cells. Upregulation can be seen for both transporters after 3 h of amino acid starvation. Upregulation of both transporters is also seen in the control cells over time. (C) Brain tissue from cortex, striatum, hypothalamus, and brainstem was taken from mice placed on 24 h starvation prior to dissection (n = 4, pooled sample). Gene expression was normalized to three housekeeping genes (Gapdh, H3a and Actb). Relative gene expression levels (±SD) of Mfsd14a were compared to normal fed controls, with gene expression of the controls set to 100%. Down regulation was seen in the hypothalamus and brainstem. (D) Relative gene expression of Mfsd14b after starvation compared to controls. Down regulation was seen in the hypothalamus. (E) Brain tissue from mice placed on high fat diet (HFD) for 8 weeks from cortex, striatum, hypothalamus and brainstem was taken (n = 4, pooled sample) and used for analysis. Relative gene expression levels (±SD) were compared to normal fed mice. Mfsd14a was upregulated in the striatum samples from mice placed on the HFD. (F) Mfsd14b was upregulated in striatum but downregulated in both hypothalamus and brainstem compared to normal fed controls. Analysis was performed with t-test and Bonferroni corrected significance levels (^∗p < 0.0493, ^∗∗p < 0.00998 and ^∗∗∗p < 0.001).