Figure 4.
Effect of RNA conformation on ANG-mediated cleavage of tRNA and 28S rRNA. (a-b) EDTA pre-treatment (20 mM) of the lysate abolishes ANG-mediated tRNA cleavage and slightly promotes 28S rRNA cleavage. (a) SYBR Gold staining and (b) Northern blotting for 5ʹ-tiRNAGly-GCC and 28S rRNA. (c-d) bRNase A-mediated digestion (10 nM) is not affected by EDTA pre-treatment. (c) SYBR Gold staining and (d) Northern blotting for 5ʹ-tiRNAGly-GCC and 28S rRNA. (e-f) Calcium chelation with EGTA (10 mM) does not inhibit ANG-mediated tRNA cleavage. (e) SYBR Gold staining and (f) Northern blotting for 5ʹ-tiRNAGly-GCC and 5ʹ-tiRNAAla-AGC