Fig. 5: Interplay of oxidative stress, ER stress, and JNK pathway in NMT1 knockdown breast cancer.

a For each group, 1 million MDA-MB-231 cells were implanted into the mammary gland fat pads of 3-week-old to 4-week-old female nude mice and tumor size was monitored weekly (left).SP600125 treatment (30 mg/kg/day) started from indicated time and lasted for two weeks. The tumor image was shown on the right. b Tumor weight from A. c Tumors from A were collected and cells were isolated from each tumor. ALDH was accessed by the ALDEFLUOR assay on viable dissociated cells. d The representative images for NMT1 and Ki67 IHC staining of tumors from A (left). Ki67 positive cells were counted. e Extreme limiting dilution analysis for the four groups in A was calculated on the website http://bioinf.wehi.edu.au/software/elda/. f ER stress related genes were knocked down via lentiviral infection in Shctrl and ShNMT1-infected SUM149 cells. Then JNK was detected by western blot. g Tumors from Fig. 2-O were collected and cells were isolated from each tumor. Then the cells were lysated and underwent western blot to detect ER stress related proteins, NMT1 and JNK. h SUM149, MDA-MB-231 and HCC1937 cells were treated with NAC (10 mM) or same volume of PBS for 48 h. Then the expression of JNK was detected by western blot. i The schematic illustration of NMT1 knockdown effects on breast cancer. Data represent the mean ± SD of 3 independent experiments where *P < 0.05, **P < 0.01 and ***P < 0.001