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. 2017 Apr 18;4:42. Originally published 2015 Feb 11. [Version 3] doi: 10.12688/f1000research.6055.3

Figure 2. Valspodar inhibits dye excluding side population in canine hemangiosarcoma cells at clinically achievable concentrations.

Figure 2.

Side population analyses were done as described in Materials and methods using cultured COSB canine hemangiosarcoma cells. ( A) Live cells were gated based on light scatter properties and exclusion of 7-AAD, and ( B) the side populations were determined based on DyeCycle Violet (DCV) efflux. Verapamil was used to inhibit ABCB1 and ABCG2 at 50–100 µM concentrations. Clinical grade and research grade valspodar was used at concentrations that were achieved in the plasma of dogs in the study (30 – 600 ng/ml) as well as at the saturating dose of 1 µg/ml. The Y-axis is DCV-blue (450+/-50 nm) emission while the X-axis is DCV-red (660 +/- 40 nm) on the LSR-II. Data were analyzed and dot plots were created in FlowJo.