Fig. 1.

Experimental validation of tRF-5s. (a) Sequence alignment of tRF-5s with their parental mature tRNAs and Northern probes. The length of each sequence and CCA, post-transcriptionally added to the 3′-end of tRNA, are indicated in parentheses and brackets, respectively. (b) Total RNA from indicated treatments in A549 cells was loaded to a denaturing polyacrylamide gel for Northern hybridization using probes indicated in panel A. Total RNA is shown for equal loading. The positions of tRF-5 and mature tRNA are indicated on the right; molecular size markers are indicated on the left. The blot was exposed for between 8 and 48 h. Data are representative of two–three independent experiments. (c) Densitometric analysis of the tRF bands was performed for Fig. 1(b), using the histogram function of Adobe Photoshop. Basically, the mean tRF intensity was normalized by the corresponding mean intensity of total RNA.