Fig. 3. Relative expression levels of ASNS to α-tubulin were dramatically increased when treating human cells with nocodazole and ASNase. Relative expression.

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Fig. 3. Relative expression levels of ASNS to α-tubulin were dramatically increased when treating human cells with nocodazole and ASNase. Relative expression levels of ASNS to α-tubulin were dramatically increased when treating human cells with nocodazole and ASNase. (A) RPE-1 cells were grown for 2 days, treated with 100 ng/ml nocodazole for 16 h and then harvested for western blot analysis. (B) RPE-1 cells were grown for 2 days, treated with 2 or 4 U/ml ASNase for 3 days and then harvested for western blot analysis. (C) CCRF-CEM and MOLT-4 cells were treated with 2 U/ml ASNase for 2 days and then harvested for western blot analysis. Experiments were independently performed at least twice. ImageJ was used to quantify all western blot data. Full blots are shown in Fig. S7. We study the relative expression ratio between ASNS and α-tubulin, under drug and no drug treatment conditions, that can be expressed as (Xn/Zn)/(Yn/Zn) which can also be equal to Xn/Yn, therefore loading control (Zn) is not required in this case [where X=band intensity of ASNS, Y=band intensity of alpha-tubulin, Z=band intensity of loading control, n=sample number]. Fold-change data are then presented. Chalongrat Noree et al. Biology Open 2018;7:bio038307 © 2018. Published by The Company of Biologists Ltd