Journal: Advanced Science
Article Title: Illuminating Mitochondrial RNA G‐Quadruplexes as Structural Brakes on RNA Granule Assembly and OXPHOS
doi: 10.1002/advs.202523462
Figure Lengend Snippet: Evaluation of MitoQUMA as a fluorescent probe for mtRNA G4s. (A) Fluorescence emission spectra of 1 µ m MitoQUMA with or without 3 µ m of different mtRNAs (mtRNA G4s: Mito152, Mito125, Mito126, Mito155, Mito3; double‐stranded mtRNA: HP18; single‐stranded mtRNA: SS12) and mtDNA G4s (Mito130, Mito27, Mito0.5‐22). (B) Fluorescence emission change of 1 µ m MitoQUMA at 630 nm vs. [mt‐nucleic acids]/[ MitoQUMA ]at λ ex = 555 nm. (C–F) Live HeLa cells were treated with 5 µ m IMT1 (C), 200 µg/mL chloramphenicol (CAPr) (D), varying concentrations of RHPS4 (E), valinomycin (10 µ m together with 200 m m KCl) or ionomycin (20 µ m together with 20 m m KCl) (F), and stained with 2 µ m MitoQUMA . (G) Confocal image of live HeLa cells with overexpression of GFP‐tagged GRSF1 were stained with 2 µ m MitoQUMA . Fluorescence intensity profiles across the yellow line in the white box were shown. (H) Live HeLa cells were transfected with siRNA to knock down GRSF1 expression, followed by staining with 2 µ m MitoQUMA . For each sample of cell image, approximately 100 cells were measured. Biological replicates ( n = 3) were taken. The data are presented as mean ± SEM, and statistical significance is determined by the two‐sided Student's unpaired t‐ test (C, D, G, and H) and one‐way ANOVA followed by Turkey's multiple‐comparison test (E and F) as (ns) not significant, ( * ) p <0.05, ( ** ) p < 0.01, and ( *** ) p < 0.001. Scale bars for cell image: 10 µm.
Article Snippet: The treatment concentrations of IMT1 (TargetMol, T8841), Chloramphenicol (TargetMol, T1205), RHPS4 (TargetMol, T6967), Valinomycin (TargetMol, TP1072), Ionomycin (Beyotime, S1672), ICG‐001 (TargetMol, T6113) and BML‐284 (TargetMol, T8820) have been provided in the figure legend.
Techniques: Fluorescence, Staining, Over Expression, Transfection, Knockdown, Expressing, Comparison
![Evaluation of MitoQUMA as a fluorescent probe for mtRNA G4s. (A) Fluorescence emission spectra of 1 µ m MitoQUMA with or without 3 µ m of different mtRNAs (mtRNA G4s: Mito152, Mito125, Mito126, Mito155, Mito3; double‐stranded mtRNA: HP18; single‐stranded mtRNA: SS12) and mtDNA G4s (Mito130, Mito27, Mito0.5‐22). (B) Fluorescence emission change of 1 µ m MitoQUMA at 630 nm vs. [mt‐nucleic acids]/[ MitoQUMA ]at λ ex = 555 nm. (C–F) Live HeLa cells were treated with 5 µ m IMT1 (C), 200 µg/mL chloramphenicol (CAPr) (D), varying concentrations of RHPS4 (E), <t>valinomycin</t> (10 µ m together with 200 m m KCl) or ionomycin (20 µ m together with 20 m m KCl) (F), and stained with 2 µ m MitoQUMA . (G) Confocal image of live HeLa cells with overexpression of GFP‐tagged GRSF1 were stained with 2 µ m MitoQUMA . Fluorescence intensity profiles across the yellow line in the white box were shown. (H) Live HeLa cells were transfected with siRNA to knock down GRSF1 expression, followed by staining with 2 µ m MitoQUMA . For each sample of cell image, approximately 100 cells were measured. Biological replicates ( n = 3) were taken. The data are presented as mean ± SEM, and statistical significance is determined by the two‐sided Student's unpaired t‐ test (C, D, G, and H) and one‐way ANOVA followed by Turkey's multiple‐comparison test (E and F) as (ns) not significant, ( * ) p <0.05, ( ** ) p < 0.01, and ( *** ) p < 0.001. Scale bars for cell image: 10 µm.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_4114/pmc13104114/pmc13104114__ADVS-13-e23462-g005.jpg)