Fig. 6.

Diagram of skin interaction with Sezary cells and effects of metformin in H9 cell line and primary SS cells. a Map of PI3K/AKT/mTORC1 pathway including members CN changes found in 43 SS individuals. Asterisks indicate therapeutic targets; b left panel, H9 cells untreated (UNT) or pre-treated for 2 h with metformin at 10 mM and then stimulated or not for 30 min with SDF-1 at 10 and 100 ng/ml were analyzed by WB for indicated proteins as showed by one representative experiment. b right panel, densitometric data normalized to β-actin were expressed as mean ± SEM of 3 independent experiments. Graph shows phospho protein levels expressed as FC respect to UNT samples *P = ≤ 0.05. c H9 cells UNT or pre-treated for 2 h with metformin at 10 mM or 1 mM or rapamycin at 30 nM were allowed to migrate in response of SDF-1 used at 300 ng/ml. d SS cells left UNT or 2 h-pretreated with metformin and then stimulated with SDF-1 at 100 ng/ml were analyzed by WB as above described as showed by one representative experiments; e SS UNT or pre-treated for 2 h with 10 mM or 1 mM of metformin or AMD3100 at 15 μM or rapamycin at 30 nM were allowed to migrate in response to SDF-1 at 300 ng/ml. In both systems, migration results are shown as an absolute number of CD4⁺ migrated cells measured by flow cytometry. Each dot represents a sample; bars represent the mean of migration counts. *P = 0.01, ° P = 0.0014; **and °° and ***; °°°P ≤ 0.05 were calculated by paired t-test