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The T m of DMPC is 24 C and to keep the temperature below the T m the immobilization was done at 20 C

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For visualization of PA localization in live cells, a Spo20 PA-binding domain 25 (MDNCSGSRRRDRLHVKLKSLRNKIHKQLHPNCRFDDATKTS) or PASS domain 67 (Addgene, 193970) fused to EGFP was cloned into pCDH-CMV-MCS-EF1-Puro