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. Author manuscript; available in PMC: 2018 Mar 5.
Published in final edited form as: J Investig Med. 2017 Nov 3;66(1):1–6. doi: 10.1136/jim-2017-000622

Figure 4.

Figure 4

Ubiquitin-specific protease 50 (USP50) reduces Ku70 protein stability. (A) Mouse lung epithelial (MLE12) cells were transfected with 0–4 µg USP50-V5 plasmids per 35 mm dish and incubated for 24 hours. Total RNA was extracted and Ku70 mRNA levels were compared by real-time qPCR. The expression of Ku70 transcripts was normalized to Gapdh. The results were also normalized to 0 µg of USP50 plasmid. (B) MLE12 cells were transfected with USP50-V5 plasmids (0–4 µg of DNA per 35 mm dish) and incubated for 24 hours. Protein expression in whole cell lysates was analyzed by immunoblotting with antibodies against Ku70, V5, and β-actin. (C) MLE12 cells were transfected with USP50-V5 plasmid and incubated for 24 hours. Cells were treated with cycloheximide (CHX) (40 µg/mL) for 0–8 hours. Expression of proteins in whole cell lysates was analyzed by immunoblotting with antibodies against Ku70, V5, and β-actin.