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To generate sgRNA-resistant STK38, site-directed mutagenesis was performed on pCDH-hygro/STK38 using the following primers: Forward: 5-TTTTATAGCAACTTGATAGCCCAGCATG-3, Reverse: 5-CATGCTGGGCTATCAAGTTGCTATAAAA-3

Research Pharmacology Drug Discovery 3, 100110

For gradual thermal denaturation, TgCBS was incubated at temperatures between 30 and 70 C for 10 min, cooled on ice for 5 min, and then residual enzymatic activity toward l-Ser was assayed in buffer containing 0.2 mM NADH, 2 M LDH, 1.5 M CBL, 0.8 mM l-Hcys, and 10 mM l-Ser

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Data were acquired using a NovoCyte Flow Cytometer (Agilent Technologies, USA)