gender: Male strain: Sprague-Dawley tissue: Skeletal Muscle treatment: W time: 01mo
Treatment protocol
Pellets were surgically implanted into the gastrocnemius muscle, and each animal received 2 pellets per limb for a total of 4 pellets per rat. Prior to use, pellets were cleaned and chemically sterilized as previously described (https://doi.org/10.1177%2F1091581814565038)
Growth protocol
Male Sprague-Dawley rats (Rattus norvegicus; age, approximately 30 d; weight, 75 to 100 g) were purchased from Envigo (Barrier 208A, Frederick, MD). Rats were allowed to acclimate in the AFRRI vivarium for at least 2 weeks prior to the start of experiments. The room was maintained at standard temperature (21 ± 2°C) and humidity (30% to 70%), with a 12:12-h light:dark cycle (lights on, 0600) and access to standard rat chow (Teklad Global Rodent Diet 8604, Envigo) and water ad libitum. Rats were pair-housed in plastic microisolation cages (23.8 × 45.4 cm) with filter tops and bedding (Teklad Sani-Chips, Envigo), changed 2 or 3 times weekly. All procedures involving animals were conducted to achieve maximal possible wellbeing of the rats, were IACUC-approved (protocol no. 2016-05-006) prior to the start of the study, and were performed in compliance with the guidelines set forth in the Guide for the Care and Use of Laboratory Animals in an AAALAC-accredited facility.
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated from gastrocnemius muscle using TRIzol® reagent (Invitrogen, Carlsbad, CA) following manufacturer’s protocol. For each metal and time point, three Affymetrix chips (Clariom S Rats) were used with 250 ng of total RNA derived from a pooled sample from 2-3 animals.
Label
biotin
Label protocol
Standard Affyemtrix protocol
Hybridization protocol
Standard Affyemtrix protocol
Scan protocol
Standard Affyemtrix protocol
Description
W2_01mo
Data processing
Raw signal intensity data was background corrected, log2 transformed and normalized with robust multi-array average (RMA) from the Affymetrix data bank using the “oligo” package in R v4.0.1 (8). Each Affymetrix chip contains 25,117 probesets, including 2163 control and unmapped probesets.
