Validated compounds from the LOPAC library inducing RBC shrinkage and vesiculation RBCs were treated with DMSO control (A), ATA (calpain inhibitor; B),

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Validated compounds from the LOPAC library inducing RBC shrinkage and vesiculation RBCs were treated with DMSO control (A), ATA (calpain inhibitor; B), tamoxifen citrate (PKC inhibitor; C), reactive blue 2 (P2Y antagonist; D), ET-18-OCH3 (PKC inhibitor; E),... Validated compounds from the LOPAC library inducing RBC shrinkage and vesiculation RBCs were treated with DMSO control (A), ATA (calpain inhibitor; B), tamoxifen citrate (PKC inhibitor; C), reactive blue 2 (P2Y antagonist; D), ET-18-OCH3 (PKC inhibitor; E), rottlerin (PKC inhibitor; F), PMA (PKC activator; G), bromoacetyl alprenolol menthane (β-AR antagonist; H) and NNC 55-0396 (T-type Ca2+ channel blocker; I). RBC shrinkage and vesiculation were measured by the significant increase in events in P2, compared with DMSO control and was observed in all conditions, with the exception of bromoacetyl alprenolol menthane (H) and NNC 55-0396 (I) which induced RBC shrinkage measured by a significant decrease RBC side scatter (J). RBCs were treated with the respective compounds for 30 min at 37°C followed by flow cytometry. All compounds were diluted in DMSO and added at 10 μM final concentration. Plots represent one of three independent measurements. Quantification of cell side scatter (SSC-A) upon DMSO, bromoacetyl alprenolol menthane and NNC 55-0396 treatment (J); results shown represent mean±S.D., n=3; *P<0.1, **P<0.01, unpaired t test was applied during the analysis. Elena B. Kostova et al. Biosci. Rep. 2015;35:e00187 ©2015 by Portland Press Ltd