Figure 4
Heterogeneity of FLIRR response with 50 µM CoCl2-treated overnight controls and upon glucose challenge (25 mM glucose/30 min) in AA PCa cells. Segmented cells were analyzed as described in Fig. 1. (A–C) The X-axis denotes segmented cell identifiers, the Y-axis the % median increase of FLIRR (NAD(P)H-a2%/FAD-a1%) from CoCl2 treated control to glucose challenge in the 3 responder categories (low, medium, high). The increases of the median ratio of mitochondrial (‘mito’) vs. whole-cell (‘wC’, without nucleus) in the same cell are compared; we are suggesting that where the increase is comparable, metabolism is driven by mitochondrial OXPHOS and where whole cell data exceeds mito, the incremental rise can be attributed to glycolysis as in cells d-2, c-9 etc. in chart A and d-3, a-10, b-3 etc. in chart (B). (D) Ranges of % median changes of FLIRR based on mitochondrial data with the number (n) of cells in each category which are merged together. (E,F) Merged FLIRR frequency distribution of the 3 response categories of CoCl2 treated controls vs. glucose challenge. (G,H) Representative FLIRR images before and after the glucose challenge, showing the rise in FLIM redox ratio.
