THs mediate proliferative effects in TCL cell lines. (A) mRNA levels of ITGAV, ITGB3, and THRA in a panel of TCL cell lines in comparison with mRNA levels in normal T cells. Results shown are the mean ± SEM of n = 3 independent experiments. (B) Protein levels of integrin αvβ3 and TRαβ obtained by flow cytometry in CUTLL1, SUDHL1, OCI-Ly12, and HuT 78 cell lines. Results are representative of n = 3 experiments. (C) CUTLL1 cells treated for 24 hours with free or agarose-bound T3 (1 nM) and T4 (100 nM) or a combination of the same concentrations of both free (TH) or AG-coupled (TH-AG) hormones were evaluated by Cell Titer Blue assay. Results shown represent the percent of proliferative cells respective to CT. (D) Cell proliferation after 24-hour treatment with TH or TH-AG compared with CT (dashed line) in the complete TCL cell line panel. (E) Evaluation of DNA synthesis by [³H]TdR incorporation of CUTLL1, HuT 78, and OCI-Ly12 cells after 24 hours of hormone treatment vs CT. (F) Cyclin mRNA expression levels in CUTLL1 cells by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) after 2-, 6-, 12-, and 18-hour treatment with TH-AG (right) and free TH (left) compared with CT. (G) Cyclin D1 mRNA expression levels in HuT 78 and OCI-Ly12 cells by qRT-PCR after a 2-, 6-, and 12-hour treatment with TH-AG and free TH compared with CT. (H) PCNA protein levels by flow cytometry in CUTLL1 cells (right) and mRNA (left) levels in CUTLL1, HuT 78, and OCI-Ly12 cells after 24-hour treatment with free TH and AG-bound TH compared with CT. qRT-PCR results shown are the mean ± SEM of at least n = 3 independent experiments. For PCNA protein levels, a representative of 4 experiments is displayed. CT, control.