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. Author manuscript; available in PMC: 2011 Mar 16.
Published in final edited form as: Oncogene. 2010 Jun 28;29(37):5193–5203. doi: 10.1038/onc.2010.257

Figure 5.

Figure 5

RNAi of HRG inhibits growth of H1047R but not E545K PI3K mutant cells. (a) Real-time qPCR analysis of HRG mRNA in control and HRG siRNA transfected MCF10A/HER2/HR cells. (b and c) IB comparing pHER3Y1289 (b) and pHER4Y1284 (c) levels in control and HRG siRNA transfected MCF10A/HER2/HR cells. (d) Association of p85 with HER3 and HER4 in control versus MCF10A/HER2/HR cells transfected with HRG siRNA (day 4 post-transfection). Cell lysates were precipitated with p85 antibody followed by IB with p85, HER3 and HER4 antibodies. (e) MCF10A/HER2/HR cell numbers on day 4, 5 and 6 following transfection with HRG siRNA. Each data point represents the mean ± SE of six replicates. (f) IB comparing pHER3Y1289 and HRG levels in control and HRG siRNA transfected MCF10A/HER2/EK and HR cells on day 4 post-transfection. (g) Determination of MCF10A/HER2/EK and HR cell numbers on day 7 after transfection with HRG siRNA. Each data point represents the mean ± SE of six replicates. NS, not significant; *, p<0.05, paired t-test. (h) MDA-MB-361 cells were transiently transfected with control or HRG siRNA. Cell numbers were measured in a Coulter counter on day 7 after transfection. Each data point represents the mean ± SE of six replicates. (i) IB comparing pAKTS473 levels in MCF10A/HER2/HR cells treated with vehicle (Ctrl), 20 μM LY294002 (LY) and 250 nM BEZ235 (BEZ) for 24 h. (j and k) Real-time qPCR comparing HRG mRNA levels in MCF10A/HER2/HR (j) and HCC1954 (k) cells treated or not (Ctrl) with LY and BEZ.