Figure 2.

Cultured CD56⁺CD127⁺ and CD127⁺ LTi-like cells maintain high RORC expression. Flow cytometry–sorted tonsil Lin⁻ cNK, CD117⁺CD56⁺CD127⁺, and CD117⁺CD127⁺ LTi-like cells were expanded ex vivo using a feeder cell mixture. (A) After expansion, RORC mRNA normalized to 18S expression. The top represents the first expansion (F1), whereas the bottom follows RORC expression in a single donor over three rounds of expansion. (B) Flow cytometry of the indicated molecules on expanded cell lines after one (F1) or three (F3) rounds of expansion. (C) Single cell clones of CD117⁺CD127⁺ LTi-like cells were generated, and RORC mRNA of clones from two donors is shown, normalized to 18S expression. Bulk cell lines from the same donors are shown in parallel. (D) Correlation between AHR and RORC mRNA from LTi-like cell clones. (E) Flow cytometry of the indicated molecules on LTi-like cell clones. Each dot in A, C, and D represents a single clone or bulk cell line. In B, a single donor representative of five is shown. In E, three representative clones are shown. The mean (horizontal bars) and SEM (error bars) are shown in A and C.