Skip to main content
. 2020 Apr 15;9(4):978. doi: 10.3390/cells9040978

Figure 6.

Figure 6

Cell-free degradation assay. (A) Protein extracts were prepared from seven-day-old seedlings of WT Col-0, abi1td, abi2 and hab1-1 mutant plants and then incubated with or without MG132, over the indicated time course. The level of recombinant ACS7 was monitored by immunoblotting, using anti-GST antibody. Actin was used as a loading control and was detected by using anti-actin antibody; (B) half-life plot for cell-free degradation of GST–ACS7 in WT Col-0, abi1td, abi2 and hab1-1 extracts; (C) cell-free degradation assay of mutated and nonmutated ACS7 in wild-type protein extracts. The level of recombinant ACS7 was monitored by immunoblotting, using anti-GST antibody. Ponceau staining was used as a loading control; (D) half-life plot for cell-free degradation of GST–ACS7, GST–ACS7 S48A, S48D, S85A and S85D mutant. ACS7 protein bands were quantified, using ImageJ software, and normalized, to the control (mock) band (set as 1).