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. 2012 Aug 29;7(8):e41127. doi: 10.1371/journal.pone.0041127

Figure 5. RPMI-8226/hRANKL/eGFP cells express hRANKL.

Figure 5

A. End-point PCR demonstrating that human RANKL mRNA is present in RPMI-8226/hRANKL/eGFP cells (labeled hRANKL). hRANKL mRNA was not detected in the corresponding eGFP-only control cell line (labeled eGFP), or in wild-type cells (labeled WT). No product was amplified in the absence of RNA (labelled no RNA) or the superscript enzyme (labeled SS-). B. Western Blot analysis detected the hRANKL monomer (∼34 kDa) in culture supernatant from RPMI-8226/hRANKL/eGFP cells - red outline. (i) Wild-type and eGFP-only expressing cell controls are included, and the rhRANKL antibody-control is also shown (∼23 kDa). (ii) Ribosomal RNA loading controls were included. C. Flow cytometric analysis of the RPMI-8226/hRANKL/eGFP-expressing cells. Representative plots showing cells stained with anti-hRANKL-PE antibody (eBioScience, 1/50 dilution) (transparent profile) and isotype control (solid profile).