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. 2018 Jul 13;9(54):30324–30339. doi: 10.18632/oncotarget.25718

Figure 4. Phosphatidylcholine-specific phospholipase C (PC-PLC) and Phospholipase D (PLD) activity in brain 72 h post-treatment presented as percent saline control.

Figure 4

(A) PC-PLC activity at 22.5 h of incubation, the peak fluorescence of the corresponding positive control in these trials, at room temperature in the dark. Cleavage of the assay substrate by PC-PLC yields a dye-labeled diacylglycerol (DAG) which fluoresces using an excitation and emission maxima of 509 nm and 516 nm, respectively. Dox administration caused a significant decrease in PC-PLC activity compared to saline treated mice (**p < 0.01). Co-administration of MESNA rescued decreased PC-PLC activity by Dox (**p < 0.01). (B) PLD activity at 1h of incubation, the peak fluorescence of the corresponding positive control in these trials, at 37° C protected from light. PLD cleaves the headgroup from phospholipids thereby releasing the choline from PtdCho. Assay reactions involving the choline produce a product that fluoresces using an excitation and emission maxima of 571 and 585 nm, respectively. Dox treatment resulted in significantly decreased PLD activity compared to saline treated controls (**p < 0.01). PLD activity in the Dox+MESNA group was not significantly different from the group receiving Dox alone.