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. 2023 Mar 2;4(4):100570. doi: 10.1016/j.xplc.2023.100570

Figure 3.

Figure 3

26S proteasome-mediated GI degradation is impaired in hos15-2 under low ambient temperature.

(A) GI protein abundance at 23°C and 16°C in the WT.

(B) GI protein levels in the hos15-2 mutant under low ambient temperatures. gi-2 35S:GI-HA and hos15-2 35S:GI-HA plants were grown for 10 days under 12L/12D conditions at 23°C and then transferred to 16°C for an additional 2 days and harvested every 4 h. The GI-HA protein levels were normalized to a Coomassie brilliant blue (CBB)-stained protein. (A and B) Data are presented as means ± SD of three independent replicates. Asterisks indicate statistically significant differences (Student’s t-test; ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05).

(C and D) GI protein abundance in gi-2 35S:GI-HA(C) and hos15-2 35S:GI-HA(D) plants at 23°C and 16°C with or without MG132. Ten-day-old plants (35S:GI-HA and hos15-2 35S:GI-HA) grown under 12L/12D at 23°C were transferred to 16°C at ZT 0 with or without MG132 (50 μM) treatment and harvested at the indicated time points. The GI-HA protein levels were normalized to HSP90. Data are presented as means ± SD of three independent replicates. Numbers below the blots are a ratio of relative GI-HA levels with and without MG132 treatment at ZT24.

(E) The relative GI-HA protein levels with and without MG132 treatment at ZT24, shown in (C) and (D), were statistically analyzed. Different letters denote statistically significant differences (ANOVA, Bonferroni post-hoc test; P < 0.05).