Supplemental data (see attachment)
Fig. S1. Flow diagram of CERISE. All ground-based preparations were performed in the laboratory on the Kennedy Space Center. The eggs of worms were prepared by alkaline bleach method with 0.5 M KOH and 1.0% NaClO after overnight incubation in M9 buffer containing 5 mg/L cholesterol at 20°C. Nine thousands (9,000) of the hatched L1 larvae in 2 ml of M9 buffer containing 5 mg/L cholesterol were packed into a monolayer polyethylene bag which was separated by U-pin. Twelve (12) ml of E. coli suspension (S-basal buffer, OD600=5) was packed into the other side of the bag as food of worms. The bags were installed in the Sample Holder made of aluminum alloy. The Sample Holders were kept at 10°C in the Glacier during transporting to the ISS by STS-129 Space Shuttle. The experiment was activated by removing the U-pin from the polyethylene bag and mixing the suspension of L1 larvae and E. coli. The Sample Holders were installed in the Measurement Experiment Units (MEUs) and then the MEUs were installed to the Cell Biology Experiment Facility (CBEF, http://iss.jaxa.jp/en/kiboexp/pm/cbef/). The worms were cultivated at 20°C for four days. After four days cultivation, the samples were microscopically observed. The experiment was terminated by freezing the bags in an onboard freezer, the Minus Eighty-Degree Laboratory Freezer for ISS (MELFI, http://www.nasa.gov/mission_pages/station/research/experiments/MELFI.html). The samples were recovered by STS-130 Space Shuttle.
Table S1. Proteins detected iTRAQ analysis with significant different expression levels of 4 days growth worms (µG vs 1G)
Table S2. Subset of muscle related genes exhibiting significant changes on DNA microarray of 4 days growth worms (µG vs 1G)
Table S3. Subset of energy metabolism related genes on DNA microarray of 4 days growth worms (µG vs 1G)
Supplemental movies
File name “4days_uG.mp4”: The movies of the space-flown worms cultured for four days under microgravity.
File name “4days_1G.mp4”: The movies of the space-flown worms cultured for four days under artificial 1G on board.
These movies were downlinked from the Japanese Experiment Module (JEM) of the ISS to the JAXA Tsukuba Control Center after four days culturing in the space environment. Magnification of the objective lens on the microscope in the JEM was 4X.
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