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atp biosensor  (Addgene inc)
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Addgene inc atp biosensor
DIMT1 and mitochondrial dysfunction in EndoC-βH1 cells. Representative immunoblots ( A ) and densitometric analyses ( B ) of mitochondrial complex protein I–V. Data are mean ± SD (n = 3). Glucose-induced (1 and 20 mM glucose) hyperpolarization of the inner mitochondrial membrane as shown by TMRM ( C ); average traces ( thick lines ) in cells treated with scramble siRNA compared to the difference in maximal hyperpolarization with FCCP in DIMT1-silenced cells ( thin lines ) are shown (scale bars: 20 μm) ( D ; data are mean ± SD; n = 3). E , <t>cytosolic</t> <t>ATP/ADP</t> ratios determined by Perceval HR ( green fluorescence ); average traces and the difference in the maximal rise in ATP/ADP ratio with oligomycin, (scale bars: 20 μm). ( F ; data are mean ± SD; n = 3). G , mitochondrial OCR is shown upon stimulation with 10 mM pyruvate as scramble cells ( black lines ) and as DIMT1-silenced cells ( gray lines ). The pyruvate-stimulated respiratory response, proton leak, ATP production and maximal mitochondrial respiration with FCCP and nonmitochondrial respiration (antimycin+rotenone) are each expressed as fold relative to basal ( H – M ; data are mean ± SD; n = 4). Statistical differences were compared by Student's t test and extreme studentized deviate test was used to determine outliers. ∗ p < 0.03 or <0.05 (as indicated), ∗∗ p < 0.01 and ∗∗∗ p < 0.001. DIMT1, dimethyladenosine transferase 1; TMRM, tetramethyl-rhodamine methyl ester.
Atp Biosensor, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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DIMT1 and mitochondrial dysfunction in EndoC-βH1 cells. Representative immunoblots ( A ) and densitometric analyses ( B ) of mitochondrial complex protein I–V. Data are mean ± SD (n = 3). Glucose-induced (1 and 20 mM glucose) hyperpolarization of the inner mitochondrial membrane as shown by TMRM ( C ); average traces ( thick lines ) in cells treated with scramble siRNA compared to the difference in maximal hyperpolarization with FCCP in DIMT1-silenced cells ( thin lines ) are shown (scale bars: 20 μm) ( D ; data are mean ± SD; n = 3). E , cytosolic ATP/ADP ratios determined by Perceval HR ( green fluorescence ); average traces and the difference in the maximal rise in ATP/ADP ratio with oligomycin, (scale bars: 20 μm). ( F ; data are mean ± SD; n = 3). G , mitochondrial OCR is shown upon stimulation with 10 mM pyruvate as scramble cells ( black lines ) and as DIMT1-silenced cells ( gray lines ). The pyruvate-stimulated respiratory response, proton leak, ATP production and maximal mitochondrial respiration with FCCP and nonmitochondrial respiration (antimycin+rotenone) are each expressed as fold relative to basal ( H – M ; data are mean ± SD; n = 4). Statistical differences were compared by Student's t test and extreme studentized deviate test was used to determine outliers. ∗ p < 0.03 or <0.05 (as indicated), ∗∗ p < 0.01 and ∗∗∗ p < 0.001. DIMT1, dimethyladenosine transferase 1; TMRM, tetramethyl-rhodamine methyl ester.

Journal: The Journal of Biological Chemistry

Article Title: Ribosomal biogenesis regulator DIMT1 controls β-cell protein synthesis, mitochondrial function, and insulin secretion

doi: 10.1016/j.jbc.2022.101692

Figure Lengend Snippet: DIMT1 and mitochondrial dysfunction in EndoC-βH1 cells. Representative immunoblots ( A ) and densitometric analyses ( B ) of mitochondrial complex protein I–V. Data are mean ± SD (n = 3). Glucose-induced (1 and 20 mM glucose) hyperpolarization of the inner mitochondrial membrane as shown by TMRM ( C ); average traces ( thick lines ) in cells treated with scramble siRNA compared to the difference in maximal hyperpolarization with FCCP in DIMT1-silenced cells ( thin lines ) are shown (scale bars: 20 μm) ( D ; data are mean ± SD; n = 3). E , cytosolic ATP/ADP ratios determined by Perceval HR ( green fluorescence ); average traces and the difference in the maximal rise in ATP/ADP ratio with oligomycin, (scale bars: 20 μm). ( F ; data are mean ± SD; n = 3). G , mitochondrial OCR is shown upon stimulation with 10 mM pyruvate as scramble cells ( black lines ) and as DIMT1-silenced cells ( gray lines ). The pyruvate-stimulated respiratory response, proton leak, ATP production and maximal mitochondrial respiration with FCCP and nonmitochondrial respiration (antimycin+rotenone) are each expressed as fold relative to basal ( H – M ; data are mean ± SD; n = 4). Statistical differences were compared by Student's t test and extreme studentized deviate test was used to determine outliers. ∗ p < 0.03 or <0.05 (as indicated), ∗∗ p < 0.01 and ∗∗∗ p < 0.001. DIMT1, dimethyladenosine transferase 1; TMRM, tetramethyl-rhodamine methyl ester.

Article Snippet: Single-cell ATP/ADP ratio measurements were carried out, using coexpression of a pericam-based ATP biosensor (Perceval HR; Addgene ID: 49083) and pHRed.

Techniques: Western Blot, Fluorescence