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. 1998 Oct 19;188(8):1421–1431. doi: 10.1084/jem.188.8.1421

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LPS-inducible neo^r gene expression from HS1,2N. Total RNA samples isolated from spleen cells of either unstimulated or day 4 or 5 of LPS activation, and were separated on a denaturing formaldehyde containing 1% agarose gel. Respectively, neo^r-, pgk-, and mb-1-RNA transcripts were detected with specific probes as mentioned in experimental procedures and are indicated with an arrow. Lanes 1, 2, and 4 contain RNA from unstimulated splenocytes that harbor a randomly integrated HS12 pgk-neo^r targeting construct (HS12RAN) and are derived from ES cell clones R12.3, R12.11 and R12.1, respectively. Lanes 7, 8, and 10, contain RNA samples of ES-derived splenocytes (again clones R12.3, R12.11 and R12.1) after 4 d of LPS stimulation, and lanes 12, 13, and 15 contain RNA samples of the same spleen cells after 5 d of LPS stimulation. Lanes 3 and 5 contain RNA samples derived from unstimulated spleen cells that harbor a targeted replacement of the HS12 pgk-neo^r targeting construct. Lanes 9 and 11 contain RNA samples of the same spleen cells after 4 d of LPS stimulation and lanes 14 and 16 contain RNA samples of the same spleen cells after 5 d of LPS stimulation. Lane 6 contains RNA from unstimulated RAG-2–deficient spleen cells.