Mouse skeletal muscle was solubilized as described above. Immunoprecipitations were then performed following the manufacturer’s instructions using the Dynabeads Protein G (Life Technologies). PLAs were performed following the manufacturer’s instructions using the Duolink anti–Mouse MINUS and anti–Rabbit PLUS In Situ PLA probes and the Duolink In Situ Detection Reagents Red (Sigma). Cells were counterstained with α-BTX–A488 for 60 min and imaged using a Zeiss LSM880 microscope with an AiryScan1 detector equipped with a 63× 1.4-NA.
Airyscan1 detector
Manufactured by Zeiss
The AiryScan1 detector is a state-of-the-art imaging device designed for confocal microscopy. It utilizes a unique detector array to improve image resolution and sensitivity compared to traditional confocal detectors.
Automatically generated - may contain errors
2 protocols using airyscan1 detector
1
Muscle Protein Interaction Analysis
2
Immunostaining of Cell Cultures and Muscle Fibers
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Incubations with primary antibodies in PBS-0.1% Tween 20 were performed either at room temperature for 60 min (C2C12 cells) or at 4°C overnight (isolated dissociated muscle fibers) and washed. After incubation for 1-3 hours at room temperature with fluorescent secondary antibodies, DNA nuclear were stained with DAPI for 10 min. Coverslips were mounted on microscope slides with FluorSave TM reagent (Calbiochem). Images were captured at RT on either a Zeiss LSM880 microscope (Carl Zeiss) with an AiryScan1 detector equipped with a preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this this version posted January 22, 2022. ; https://doi.org/10.1101/2022.01.21.477182 doi: bioRxiv preprint Whitney) was applied. Data distribution was assumed to be normal, but this was not formally tested. For a multiple factorial analysis of variance, two-way ANOVA was applied. P-values under 0.05 were considered statistically significant (shown as a single asterisk in figures); pvalues under 0.01 were considered highly statistically significant (shown as two asterisks in figures); p-values under 0.001 were considered very highly statistically significant (shown as three asterisks in figures).
The copyright holder for this this version posted January 22, 2022. ; https://doi.org/10.1101/2022.01.21.477182 doi: bioRxiv preprint Whitney) was applied. Data distribution was assumed to be normal, but this was not formally tested. For a multiple factorial analysis of variance, two-way ANOVA was applied. P-values under 0.05 were considered statistically significant (shown as a single asterisk in figures); pvalues under 0.01 were considered highly statistically significant (shown as two asterisks in figures); p-values under 0.001 were considered very highly statistically significant (shown as three asterisks in figures).
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