FIGURE 9.

Complex amino acid requirements for full activation of mTORC1 in oocytes. Un-injected oocytes were incubated for 4 days in OR²⁺ (pH 7.8) buffer. In the experiment, oocytes were incubated with different amino acid mixtures (1 mM each) for 4 h. Activation of mTORC1 was measured by immunodetection of ribosomal protein S6 and phospho-S6. Densitometry in each bar graph was normalized to the negative control, where oocytes were incubated without amino acids in 1 $\times$ ND96 buffer only (last column). Bar graphs represent the mean ± S.E.M. of 3 experimental repeats (e = 3) with each condition in each repeat representing the protein expression of 12 oocytes (n = 12). The amino acid composition of each group shown below the densitometry bar graphs was based on several criteria: substrate affinity for SNAT2, previously shown ability to activate mTORC1, and amino acids previously shown to activate mTORC1 when injected into oocytes (Christie et al., 2002). Amino acid groups are indicated by numbers 1, 2, 3, 4, 5 in the margin. The graph was divided into panels A and B for clarity.