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For VAPB WT versus P56S experiments, cells were co-transfected with 40 ng proinsulin-NLuc and 200 ng of either pEGFP-VAPB, pEGFP-VAPB-P56S [generated in house by site directed mutagenesis from pEGFP-VAPB with Fwd 5-CGATGATTCCGCTGTTGCTCCTCACACAGTACCTAC-3 and Rev 5- GTAGGTACTGTGTGAGGAGCAACAGCGGAATCATCG-3 mutagenesis primers and the Quick-Change Lightning kit (Agilent)], or pcDNA3.1+ as a control

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The primary issue is that meds are replacing exercise and diet, resulting in more muscle loss, argues one rheumatology and internal medicine physician on Sermo

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