Retroviral expression of RXRα constructs. (A) Schematic representation of full-length wild-type RXRα and truncated RXRα (RXRαΔ) containing the ligand binding domain (LBD) lacking the 5′ AF-1 and DNA-binding domains (DBD). cDNAs were inserted into a retroviral backbone 5′ of an IRES-GFP cassette. (B) Western blot analysis of untransduced (WT) or gene-transduced U937Te cells. GFP⁺ cells were sorted prior to analysis (CTRL indicates empty control vector; RXRαΔ- or RXRα-encoding vectors). Protein extracts were prepared using cell lysis buffer (“Materials and methods”). (C) U937Te cells were transduced with RXRαΔ or empty control vector. After transduction (48 hours), cells were stimulated with VD3 (60 nM) for 48 hours and were then analyzed for GFP versus CD11b and CD14 surface expressions. GFP^hi, GFP^lo, and GFP⁻ cells were separately gated and analyzed for CD11b versus CD14. (D) Representative FACS analysis of CD54 of gene-transduced GFP^hi HL60e cells. HL60e cells were gene transduced with RXRαΔ or empty control vector (CTRL). After transduction (48 h), cells were stimulated with 9cRA (100 nM), ATRA (100 nM), or vehicle (0.001% DMSO). Data in panel B are representative of 2 experiments. Data in panels C and D are representative of 5 experiments.