Figure 6
Immediate effects of isotretinoin on Treg suppression/Teff proliferation.
Proliferation of Teffs was assessed by FACS analysis by measuring the dilution of the intensity of CFSE. (A) Proliferation of Teffs for the control group (top) and isotretinoin group (bottom) at basal conditions. (B) Unlabeled Teffs co-cultured with CFSE-labeled Teffs from vehicle treated animals at a 1/2 or 1/8 ratio (top) or Tregs co-cultured with Teffs from the controls at a 1/2 or 1/8 ratio (bottom). (C) Unlabeled Teffs co-cultured with CFSE-labeled Teffs from isotretinoin treated animals at a 1/2 or 1/8 ratio (top) or Tregs co-cultured with Teffs from the controls at a 1/2 or 1/8 ratio (bottom). (D) Inverse setting: unlabeled Teffs from controls were cultured with CFSE-labeled Teffs from isotretinoin treated animals at a 1/2 or 1/8 ratio or Tregs from controls cultured with Teffs from the isotretinoin condition. (E) Inverse setting: unlabeled Teffs from isotretinoin treated animals were cultured with CFSE-labeled Teffs from control animals at a 1/2 or 1/8 ratio or Tregs from isotretinoin treated animals were cultured with Teffs from controls. Shown are representative graphs of two independent experiments. Solid line: stimulated CFSE-labeled Teffs. Dashed line: unstimulated CFSE-labeled Teffs. Abbreviations: Teffs, effector T-cells (CD4+CD25−); Tregs, regulatory T-cells (CD4+CD25+); unlab, unlabeled Teffs; ctrl, vehicle treated animals; iso, isotretinoin treated animals; 1/2 and 1/8 ratio of unlab or Tregs; FACS, fluorescence-activated cell sorting; CFSE, carboxyfluoresceinsuccinimidyl ester.
