Cytofluorometric analysis of ES-D3 clones. (A) Untransfected ES-D3 cells or ES-D3 cells transfected with pXT1-Ptpγ full-length cDNA in the sense orientation (clones S5 and S9) were analyzed: black, S9 without primary antibody (result was the same for parental and S5); red, S9 with irrelevant rabbit IgG (result was the same for parental and S5 cell lines); blue, parental cells reacted with anti–Ptpγ P4 antibody; green and purple, the individual S5 and S9 clones expressing Ptpγ RNA, reacted with anti–Ptpγ P4 antibody. The signal for the empty vector transfected clones was the same as for the parental ones and has been omitted in the figure. (B) S5 and S9 PTPγ overexpressing clones were supertransfected with either antisense construct B (ASpXT1S5 and AS6pXT1S9 clones) or sense construct B ( S4pXT1S5 and S8pXT1S9 clones) and the resultant stable clones tested for expression of the exogenous Ptpγ as in (A): black, S9 without primary antibody; red, antisense B supertransfectant clone AS6pXT1S5 reacted with anti–Ptpγ P4 antibody; blue, antisense B supertransfectant, AS6pXT1S9, reacted with anti–Ptpγ P4 antibody; green and purple, two sense B supertransfectants reacted with anti–Ptpγ P4 antibody. The B antisense construct supertransfectants have down-modulated surface expression of the exogenous Ptpγ in the S5 and S9 clones. Both panels represent one of two independent experiments that gave the same results.