Fig. 5
ACTN4 knockdown exerted no additive anti-cancer and anti-metastatic effects induced by EA on breast cancer. a MDA-MB-231 and (b) BT-549 cells were transfected with siACTN4 for 48 h and treated with or without 25 μM EA for additional 24 h. LY294002 (10 μM) was used as a positive control substance for this assay. The findings revealed that siACTN4 did not aggravate the inhibition effects of EA on breast cancer cells when used in combination; c-d siACTN4 did not significantly aggravate the inhibition effects of EA on cell migration and invasion of MDA-MB-231 cells. LY294002 (10 μM) was used as a positive control substance for this assay; e For xenograft models, 106 cells of MDA-MB-231 with or without ACTN4 knockdown were subcutaneously implanted into the mammary fat pads of female NOD/SCID mice at 4-week-old in the presence or absence of EA (50 mg/kg/d). There was no additive inhibitory effects on tumor growth when combining EA with ACTN4 knockdown; f The ALDEFLUOR assay showed that the ALDH+ subpopulations in the tumor essence between EA alone and EA plus shACTN4 groups