Fig 2: Increased expression of MRP1 in sarcoma spheroids. Real-time PCR examination of MRP1 transcripts in the 2D monolayers (“2D”) and 3D spheroids (“3D”) of MG-63 (A) and SW-1353 (D; n = 4 for each). Expression levels are shown as a ratio compared with ACTB. Protein expression of MRP1 in the lipid raft-enriched protein lysates of both groups. Blots were probed with anti-MRP1 (approximately 250 kDa) and anti-ACTB (43 kDa) antibodies (B, E). Summarized results were obtained as the optical density of MRP1 (C, F) band signals. After compensation for the optical density of the MRP1 protein band signal with that of the ACTB signal, optical density in “2D” was expressed as 1.0 (n = 4 for each). Results are expressed as means ± SEM. *, **P < .05, .01 vs 2D

Fig 3: Decreased expression of MRP1 by the inhibition of KCa1.1 in sarcoma spheroids. Real-time PCR examination of MRP1 transcripts in vehicle- and PAX-treated MG-63 (A) and SW-1353 (D) spheroids (n = 4 for each). Expression levels are shown as a ratio compared with ACTB. Protein expression of MRP1 in the lipid raft-enriched protein lysates of both groups. Blots were probed with anti-MRP1 (approximately 250 kDa) and anti-ACTB (43 kDa) antibodies (B, E). Summarized results were obtained as the optical density of MRP1 (C, F) band signals. After compensation for the optical density of the MRP1 protein band signal with that of the ACTB signal, optical density in the vehicle control was expressed as 1.0 (n = 4 for each). Results are expressed as means ± SEM. **P < .01 vs vehicle control