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. 2020 Feb 21;32(4):1270–1284. doi: 10.1105/tpc.19.00791

Figure 9.

Figure 9.

Down-regulating KETCH1 Reduced Ribosomal Biogenesis and Translational Efficiency.

(A) Polysome profiling assay with sucrose density gradient. The OD260 absorption (A260) was monitored together with fractionation. The fractions containing 40S, 80S of ribosome, and polysomes in wild type (WT) or Pro35S:amiR-KETCH1 (amiR) are indicated. Three biological replicates were performed, and similar results were obtained.

(B) CLSM of a GFP-translational fusion of ARF2 or ARF3 from wild-type or Pro35S:amiR-KETCH1 protoplasts transformed with ProUBQ10:uORF-ARF2-GFP or ProUBQ10:uORF-ARF3-GFP. Dotted lines illustrate protoplast silhouettes and arrowheads point at the nuclei.

(C) and (D) GFP intensity in the nucleus for ARF2 (C) or ARF3 (D) from wild-type or Pro35S:amiR-KETCH1 protoplasts transformed with ProUBQ10:uORF-ARF2-GFP or ProUBQ10:uORF-ARF3-GFP. a.u., Arbitrary fluorescence unit. Results are means ± SD (n > 28) from two batches of transformation events. Asterisks indicate significant difference (t test, P < 0.05). Bars = 10 µm.