PI/RNase Staining Buffer

Protocol tips

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	- Cells were fixed at −20 °C in 70% ice cold ethanol for at least 12–18 h and was done at different intervals.

Protocol tips
- Cells were fixed at −20 °C in 70% ice cold ethanol for at least 12–18 h and was done at different intervals.

Publication protocol

Cells were synchronized at G0/G1 by addition of 1 mM hydroxyurea (Sigma, http://www.sigmaaldrich.com/catalog/product/sigma/h8627?lang=fr®ion=CH), an inhibitor of DNA synthesis, for 3 h. Cells were then washed twice with PBS and cultured with fresh GM to resume cycling into S and G2/M phases. Cells were accumulated within S phase 2 h afterwards, followed by G2/M phase accumulation 6 h after the release. Cells were fixed at various intervals of growth under different gravitational conditions and stored at −20 °C in 70% ice cold ethanol for at least 12–18 h prior to staining. Cell cycle was analyzed based on DNA content through propidium iodide (PI)/RNase staining (BD Biosciences, catalog number 550825, http://www.bdbiosciences.com/ptProduct.jsp?ccn=550825) by FACS

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