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(A, C) Immunofluorescence of cells treated for 5 h with EtBr, water, IMT1B, or DMSO. TOM20 was used as a mitochondrial marker in all panels. TOM20 is shown in magenta in the MERGE panel and GRSF1 in cyan. (B, D) Immunofluorescence of cells treated for 5 h with EtBr, water, IMT1B, or DMSO. TOM20 was used as a mitochondrial marker in all panels. TOM20 is shown in magenta in the MERGE panel and <t>FASTKD2</t> in cyan. DAPI is shown in gray. Boxes in MERGE panels represent areas enlarged. Zoom-ins show RNA staining in gray. Arrowheads mark mitochondrial RNA granules. Scale bars in whole-cell panels represent 10 μm and in zoom-ins represent 2 μm.
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(A, C) Immunofluorescence of cells treated for 5 h with EtBr, water, IMT1B, or DMSO. TOM20 was used as a mitochondrial marker in all panels. TOM20 is shown in magenta in the MERGE panel and GRSF1 in cyan. (B, D) Immunofluorescence of cells treated for 5 h with EtBr, water, IMT1B, or DMSO. TOM20 was used as a mitochondrial marker in all panels. TOM20 is shown in magenta in the MERGE panel and FASTKD2 in cyan. DAPI is shown in gray. Boxes in MERGE panels represent areas enlarged. Zoom-ins show RNA staining in gray. Arrowheads mark mitochondrial RNA granules. Scale bars in whole-cell panels represent 10 μm and in zoom-ins represent 2 μm.

Journal: Life Science Alliance

Article Title: Transcription arrest induces formation of RNA granules in mitochondria

doi: 10.26508/lsa.202403082

Figure Lengend Snippet: (A, C) Immunofluorescence of cells treated for 5 h with EtBr, water, IMT1B, or DMSO. TOM20 was used as a mitochondrial marker in all panels. TOM20 is shown in magenta in the MERGE panel and GRSF1 in cyan. (B, D) Immunofluorescence of cells treated for 5 h with EtBr, water, IMT1B, or DMSO. TOM20 was used as a mitochondrial marker in all panels. TOM20 is shown in magenta in the MERGE panel and FASTKD2 in cyan. DAPI is shown in gray. Boxes in MERGE panels represent areas enlarged. Zoom-ins show RNA staining in gray. Arrowheads mark mitochondrial RNA granules. Scale bars in whole-cell panels represent 10 μm and in zoom-ins represent 2 μm.

Article Snippet: All antibodies were diluted in blocking buffer, if not stated otherwise: TOM20 (#sc-17764; Santa Cruz) used in a 1:200 to 1:500 dilution, GRSF1 (#ab205531; Abcam) used in a 1:500 to 1:1,000 dilution, FASTKD2 (#17464-1-AP; ProteinTech) used in a 1:500 dilution, ds/ssDNA (#61014; PROGEN) used in a 1:100 dilution, LRPPRC/GP130 (#ab97505; Abcam) used in a 1:200 dilution, SUV3L (#ab127909; Abcam) used in a 1:1,000 dilution in milk for Western blot, ACTB (#3700; Cell Signaling) used in a 1:5,000 dilution in milk for Western blots, HRP-conjugated anti-rabbit IgG (cat. no. 7074S; Cell Signaling) used in a 1:10,000 dilution in 5% milk, HRP-conjugated anti-mouse IgG (cat. no. 70765; Cell Signaling) used in a 1:10,000 dilution in 5% milk, goat anti-rabbit IgG Alexa Fluor 647 (cat. no. A-21245; Thermo Fisher Scientific) used in a 1:1,000 dilution, goat anti-mouse IgG2a Alexa Fluor 488 (cat. no. A-21131; Thermo Fisher Scientific) used in a 1:1,000 dilution, and goat anti-mouse IgM Alexa Fluor 555 (cat. no. A-21426; Thermo Fisher Scientific) used in a 1:1,000 dilution.

Techniques: Immunofluorescence, Marker, Staining