Immuno-Northern Blotting: Detection of RNA Modifications by Using Antibodies against Modified Nucleosides
Fig 5
Detection of stress-induced tRNA-derived fragments by immuno-northern blotting.
(A) The image of stress-induced tRNA cleavage. tRNAs are cleaved under conditions of cell stress, and consequently, tRNA-derived fragments such as the 5’-side and 3’-side fragments appear. The modification of m1A is usually contained in the 3’ side of the tRNA structure. (B) Total RNA (0.75 μg) was isolated from HK-2 cells treated with sodium arsenite (500 μM) for the indicated periods and analyzed by SYBR staining and INB with anti-m1A antibody using acrylamide gel separation. Arsenite-stress induced tRNA cleavage, and the resultant tRNA-derived fragments were detected by SYBR staining and firmly detected by the INB with anti-m1A antibody.