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Fig. 2 | Molecular Cancer

Fig. 2

From: Posttranscriptional upregulation of HER3 by HER2 mRNA induces trastuzumab resistance in breast cancer

Fig. 2

The HER2 3’UTR regulates HER3 expression. (A) Microarray heat map of differentially expressed genes in the ErbB pathway in T47D cells transfected with the HER2 3’UTR and control vector. See also Tables S1 and S2. (B) Real-time PCR analysis of gene expression in T47D cells as in (A). ***, P < 0.001 compared to the control. (C, D) Real-time PCR (C) and western blotting (D) analysis of HER3 mRNA and protein levels in T47D and MCF7 cells transfected with the HER2 3’UTR and control vector. (E) Real-time PCR analysis of HER3 expression in breast primary tumor cells transfected with the HER2 3’UTR and control vector. (F) FACS analysis of HER2 and HER3 expression in T47D and MCF7 cells stably transfected with the HER2 3’UTR or control vector. (G) Immunostaining of HER3 levels in T47D and MCF7 xenograft tumors. Red, HER3; Blue, DAPI. Scale bar, 40 μm. The mean values of HER3 fluorescence calculated and analyzed by Leica TCS SP8 Confocal platform are shown. (H) Western blot analysis of the PI3K/AKT pathway of T47D cells transfected with the HER2 3’UTR or control vector. (I, J, K) Western blot (I, K) and real-time PCR (J) analysis of EGFR, HER2, HER3 and HER4 protein and HER3 mRNA levels in T47D cells transfected with control vector, the HER2 3’UTR, or the HER2 CDS. All error bars represent the standard deviation. All quantitative data were generated from a minimum of three replicates

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