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. 2022 Apr 21;33(5):ar44. doi: 10.1091/mbc.E21-12-0611

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FIGURE 4: — Testing the functionality of the central core of the TOR2 MAS domain. (A) TOR2 MAS domain plasmid pPL626 containing the S1972R (RapR) TOR1-1 allele was modified to create the indicated TOR1-TOR2 chimeras. Amino acid numbering refers to TOR2 protein sequence. “N” and “C” refer to N- and C-termini, respectively. Plasmids were introduced into heterozygous TOR2/tor2Δ cells and analyzed by tetrad analysis as described in the legend to Figure 3A. (B) Western blot analysis of TOR1-TOR2 chimeras. TOR2/tor2Δ cells carrying the indicated plasmids were grown in selective media to mid–log phase, and protein extracts were prepared and analyzed by SDS–PAGE and immunoblotting, as described in the legend to Figure 2D. (C) TOR2/tor2Δ cells carrying the indicated plasmids were tested for rapamycin resistance as described in the legend to Figure 3B.