Fig. 5.

Naringenin (Nar) inhibits TGF-β1 secretion and the invasion of 4T1 cells in vitro. a Invasion of cultured 4T1/RFP cells (upper) and 4T1/TGF-β1 cells (lower) after treatment with 100 μM of Nar or 5 μg/ml of 1D11 for 48 hours. The transwell cells were stained and imaged under microscopy (100×) following fixation (left). The stained cells were dissolved using DMSO and OD590 values were determined using the multimode reader (right). b Secreted TGF-β1 concentration in supernatant from 4T1/RFP cells and 4T1/TGF-β1 cells after treatment with Nar or 1D11 for 48 hours. c Flow cytometry analysis of the percentage of Tregs induced by the cell supernatants. Percentage gated cells are shown (upper). Column charts show the mean percentage obtained from three experiments (lower). d TGF-β1 concentration in 4T1/RFP cells and 4T1/TGF-β1 cells after treatment with Nar or 1D11 for 48 hours. *P <0.05, **P <0.01, ***P <0.001. Error bars indicate SE. TGF transforming growth factor, Treg regulatory T cell