A Tissue Retrieval and Postharvest Processing Regimen for Rodent Reproductive Tissues Compatible with Long-Term Storage on the International Space Station and Postflight Biospecimen Sharing Program

Figure 6

Effect of different extraction and storage methods on RNA and protein quality in ovaries, and testes extracted and/or stabilized in TRIzol or RNAlater. (a) Total RNA integrity analysis. Total RNA was isolated from mouse testis and stabilized in TRIzol reagent (RNA yield and quality control (lane 1); RNAlater for 2 wks at 4^°C (lane 2); RNAlater for 1 wk at room temperature and 1 wk at 4^°C (lane 3); RNAlater for 2 wks at RT (lane 4). (b) Agarose gel electrophoresis of PCR products with primers for IL-1α and GAPDH. For RT PCR total RNA was used after stabilization in TRIzol reagent (lane 1); RNAlater for 2 wks at 4^°C (lane 2); RNAlater for 1 wk at RT and 1 wk at 4^°C (lane 3); RNAlater for 2 wks at RT (lane 4). (c) Total RNA integrity analysis STS-131 ground (G10, G11, and G12) and flight (F10, F11, and F12) uteri fixed in RNAlater after 30–40 min after euthanasia. (d) Comparison of buffers to remove RNAlater for subsequent western analysis of β-actin integrity in ovaries. Mouse ovaries were stored at RNAlater for 1 wk at RT or 2 wk at 4^°C; tissues were homogenized in RIPA lysis buffer (lanes 1, 2) or ProteoJET Lysis reagent (LR) (lanes 3, 4). Total cell lysates were prepared and subjected to SDS-PAGE. Western for β-actin is presented.