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. 2022 Jun 14;50(12):6919–6937. doi: 10.1093/nar/gkac495

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© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — (A) Northern blotting of total RNA (1.5 μg) extracted from HeLa cells after time-limited exposure (1 h) to As[III] (0.5 mM), followed by culturing cells for the indicated times (1, 2, 4, 6, 8 h) after the removal of As[III], using probes against the 5′ ends of tRNA-Gly^GCC/CCC and tRNA-Ala^AGC. Black arrowhead: mature tRNAs; red arrowhead: tsRNAs; asterisks: digitally enhanced against parental tRNA signals. (B) Northern blotting of total RNA (3 μg) extracted from HeLa, U2OS, HEK293 cells and i-MEF, which had been exposed (1 h) to increasing molarities of As[III], followed by culturing cells for 24 or 48 h after the removal of As[III] using a probe against the 5′ end of tRNA-Gly^GCC/CCC. Black arrowhead: mature tRNAs; red arrowhead: tsRNAs; asterisks: digitally enhanced against parental tRNA signals. (C) Indirect immunofluorescence image of HeLa cells after time-limited exposure (1 h) to As[III] (0.5 mM), followed by culturing cells for 24 h after the removal of As[III], using antibodies against G3BP1 (magenta) and TIA-1 (cyan). Individual insets: DNA (black). Scale bar 10 μm. (D) Indirect immunofluorescence image of HeLa cells after exposure (4 h) to staurosporine (1 μM), using antibodies against cleaved caspase 3 (cyan) as indicator of apoptotic cells and phosphorylated γ-H2AX (magenta) as indicator of DNA damage. DNA: black. (E) Northern blotting of total RNA (1.5 μg) extracted from HeLa cells, which had been exposed to staurosporine (1 μM) for the indicated times (1, 2, 4, 6 h) using probes against the 5′ ends of tRNA-Gly^GCC/CCC and tRNA-Ala^AGC. Black arrowhead: mature tRNAs; red arrowhead: tsRNAs; asterisks: digitally enhanced against parental tRNA signals.