Conditional activity and selectivity of the APD-CLDs

Published: 04-08-2018| Version 1 | DOI: 10.17632/9knckkzfg7.1
Kristian Mark Jacobsen,
Nikolaj Villadsen,
Thomas Tørring,
Michail Tsakos,
Thomas B. Poulsen


Figure 1b: Viability of PANC-1 cells treated with 1 or 2a for 48 hours under normoxia (red) or hypoxia (blue). Viability was assessed with CellTiter BlueTM (Promega). Figure 1d/Figure S1a: Normoxic treatment of PANC-1 cells with 2b for 8 or 24 hours followed by 24 hours of hypoxic incubation confirms the stability of the APD-CLDs in normoxic cell culture. N = normoxia, H = hypoxia. Viability was assessed as in (b). Figure 1e/Figure S1d: PANC-1 cells were treated with 2b under normoxia for 4.5, 8 or 24 hours before the medium was exchanged for fresh complete medium and cells were placed under hypoxia for additionally 24 hours. For comparison cells were kept under normoxia for 24 hours followed by 24 hours under hypoxia in presence of 2b (No wash). Figure S1c: PANC-1 cells were pretreated with hypoxia or normoxia for 47 hours before they were treated with rakicidin A under normoxia for 24 hours. N = 3. Figure S1f: Stability of rakicidin A (20 µM) in H2O/MeCN (1:1) containing either N-acetylcysteine (5 mM) and N-acetylglycine (5 mM). Injections of the mixtures were performed on a HPLC every third hour over the course of 1 day on a C18 column (Kinetex, 4.6 x 250 mm, C18, 5 µm, 100 Å, Phenomenex) equipped with a guard column and eluted with MeCN/H2O (40:60 --> 100:0 over 33 minutes). Retention time = 20-21 minutes. The absorbance of the APD chromophore was followed at 262 nm. N = 1.