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To this end, we synthesized a number of permethylated quinolinium analogues[17] guided by in silico calculation of partition coefficient (clogP).[26] Addition of hydrophobic methyl group substitutions around the quinolinium scaffold (previously shown to negatively impact NNMT inhibitory activity)[17] only slightly improved membrane permeability via passive transport as indicated by the low, but non-zero, permeability values for 1,8-diMQ and 1,2,4,8-tetraMQ (Table 1)

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The cells were then washed using MACS buffer (0.5% BSA + 2mM EDTA in PBS) followed by staining with different clones of anti-CD38: LS198-4-2 (Beckman Coulter), JK36 (Beckman Coulter) and HIT2 (BD Biosciences) for 10 mins in the dark at room temperature