Figure 6

From: Detecting intratumoral heterogeneity of EGFR activity by liposome-based in vivo transfection of a fluorescent biosensor

Figure 6

Effect of nitric oxide inhibitor on EGFR activity in HCC1954 cells co-cultured with activated macrophages. (a) mRNA level of iNOS was detected by qPCR (relative to the house keeping gene Tata-binding protein) in mouse macrophages treated with mouse tumour necrosis factor alpha (50 ng/ml) or conditioned media of HCC1954 cells for 3 days. One-way ANOVA to test differences between group means. P values are derived from a post-hoc Tukey’s test subsequently performed to compare means of individual groups while controlling for family-wise error rate. The difference was statistically significant between control and treatment groups, data were expressed as means±s.e.m. (*P<0.005, N=3). (b) HCC1954 cells expressing Picchu-X biosensor were co-cultured with activated macrophages from (a) for 48 h. Treated with L-NAME (2 mM) for 1 h, fixed and imaged. Presence of macrophages significantly increased FRET efficiency (**P<0.0001, N=6), which was significantly decreased by iNOS inhibitor (*P=0.01). (c) FRET/FLIM images of cells with and without L-NAME; and with and without activated macrophages. Scale bar is 25 μm.