Fig. 4: miR-200b/c and their target IKKβ induce NF-κB activation in stromal fibroblasts | Cell Death & Disease

Fig. 4: miR-200b/c and their target IKKβ induce NF-κB activation in stromal fibroblasts

From: Primed atypical ductal hyperplasia-associated fibroblasts promote cell growth and polarity changes of transformed epithelium-like breast cancer MCF-7 cells via miR-200b/c-IKKβ signaling

Fig. 4

a Luciferase assay was used to test relative luciferase activities of IKKβ in 293T cells co-transfected with the indicated miR-200b/c or control vector with corresponding reports. b Western blotting analysis to check the protein levels of IKKβ, IκBα, p-IκBα, and nuclear P65 in the indicated engineered CAFs or NFs. GAPDH or histone 3 (H3) was used as a loading control. c Immunofluorescence staining of nuclear translocation of P65 in the indicated engineered CAFs or NFs (magnification ×200). d ELISA-based measurement of P65 activity to show NF-κB function in the indicated engineered CAFs or NFs. e Western blotting analysis of IKKβ, IκBα, p-IκBα, and nuclear P65 protein expressions in the NFs, NFs/IKKβ with or without miR-200b/c encoding vector. GAPDH or histone 3 (H3) was used as a loading control. f Immunofluorescence staining to show nuclear translocation of P65 in the NFs, NFs/IKKβ with or without miR-200b/c expression vector (magnification ×200). g ELISA-based measurement of P65 activity to detect NF-κB function in the NFs, NFs/IKKβ with or without miR-200b/c. The data were shown as mean ± SD for N ≥ 3 separate experiments, *p < 0.05