Figure 4
From: Activation of PKC supports the anticancer activity of tigilanol tiglate and related epoxytiglianes
PKC isoform activation profile and ERK phosphorylation in response to epoxytigliane analogues. (A) Examples of PKCα, − βII, − γ, − δ and − θ translocation after treatment with vehicle (Vehc.) or 500 nM TT (EBC-46), EBC-83 or PMA. HeLa cells transfected with PKC-EGFP isoforms were incubated with the indicated concentrations of epoxytigliane for 1 h. Images were acquired using a GE InCell Analyzer 2000. These images were also used for the quantitation shown in (B). (B) Heatmap depicting PKC isoform translocation profile (− α, − βI, − βII, − γ, − δ, − θ, − η and − ζ—mean % of cells showing EGFP translocation to the plasma membrane) in response to 500, 50 and 5 nM TT (EBC-46) and the indicated analogues. > 150 cells counted per biological replicate. n = 3. (C) Analysis of ERK phosphorylation in response to administration of TT (EBC-46) and the indicated analogues. Cell extracts were analyzed using single plex phospho-ERK and total ERK alphaLISA kits from MM649 cells. Data expressed in box plot format (min to max values) using mean fluorescence intensity values (MFI) from phospho-ERK experiments normalized to MFI values from total ERK (p-ERKF/ERKF). n = 4.
