ZymoBIOMICS RNA Miniprep Kit

Protocol tips

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	Total RNA isolation was performed with RNA MiniPrep kit (Zymo research). cDNA synthesis was carried out as previously described by Gilkes et al.

Protocol tips
Total RNA isolation was performed with RNA MiniPrep kit (Zymo research). cDNA synthesis was carried out as previously described by Gilkes et al.

Publication protocol

Matrix embedded cells with cell densities of 10 cells mm−3 and 50 cells mm−3 were incubated for 24 h at 37 °C in a humidified incubator. The matrices were exposed to cell lysis buffer and mechanically broken down using a syringe. The suspension was centrifuged and the supernatant was measured for STAT3 and PhosphoSTAT3 using an ELISA kit (Abcam). WASF3 expression was measured using qRT-PCR for the cell densities stated previously and matrix embedded cells with a cell density of 10 cells mm−3 exposed to IL-6 and IL-8 alone and in combination at the precise concentrations found at a high cell density of 50 cells mm−3.

Total RNA isolation was performed with RNA MiniPrep kit (Zymo research). cDNA synthesis was carried out as previously described by Gilkes et al.

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