Muscle

Adult male thoraxes were prepared for fixation by removing the head and abdomen. Additionally, the tips of the legs were cut using micro-scissors to allow easy access to the leg muscles for the fixative. The dissected samples were fixed in 4% paraformaldehyde for 30 min. The samples were subsequently embedded in 4% low-melt agarose and left at 4°C over night for the agarose to solidify. Samples in agarose blocks were mounted on to the stage of a vibratome (Leica VT1000M) in ice-cold PBS and 100 μm sections were cut. The sections were further trimmed under the microscope tissue sections with some surrounding agarose were transferred to 2 ml tubes. Subsequent immuno-staining steps were similar to as discussed in the previous section. Stained sections were mounted in Vectashield mounting media and imaged using a Zeiss LSM 780 confocal microscope.