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. 2004 Aug 19;32(15):4448–4461. doi: 10.1093/nar/gkh748

Figure 6.

Figure 6

Fractionation of total tRNAs isolated from CHO L1 and Cos1 L1 cells with or without transfection of the tRNASer(CGA) gene and northern blot analysis for aminoacyl (aa) forms of methionine tRNA. (A) Total tRNAs on 10% denatured PAGE, extra tRNA bands induced by tRNASer(CGA) expression construct are marked by solid arrows on the right side. (B) Analysis of uncharged and aa-forms of methionine tRNA. Parts of the tRNA samples denoted by an asterisk was treated with 0.5 M Tris at pH 9.0 to strip aa-tRNAs before fractionated on the denaturing PAGE as described in Materials and methods. tRNA blots were probed with a 32P-lableled oligonucleotide complementary to specific regions of tRNAMet(ATG). Uncharged and aminoacyl forms of methionine tRNA are marked on the left position. The relative levels of uncharged and aminoacyl-methionine tRNA shown by arbitrary units (number) were measured from the blots by densitometry. The aa-methionine tRNA signals are shown by the arbitrary units in parentheses.