Fig. 5
From: Multiplexed activity metabolomics for isolation of filipin macrolides from a hypogean actinomycete
Filipin-induced death is cCAS3 independent and pendant alkyl chain dependent. (a) Single cell contour plots of apoptotic marker cCAS3 and Ax700 viability stain. Cells were treated for indicated time then stained with Ax700, fixed, permeabilized, and stained for cCAS3. Compounds 1, 2, and 4 disrupted cell membranes without activating cCAS3. (b) Cells were challenged with each filipin and stained with propidium iodide (cell viability, DNA) and quinacrine (intracellular ATP) cocktail. Compounds 1, 2, and 4 exhibit robust PI labeling (white arrows) and concomitant loss of ATP. Compound 3 does not enhance PI infiltration of loss of ATP (white arrowhead). (c) Quantification of percent positive PI or quinacrine positive cells versus vehicle control. Cells were challenged with 10 μM of each compound for the indicated time. *p < 0.05 vs vehicle control. Scale bar = 25 µm; Red, PI = DNA; Green, quinacrine = ATP
