Fig. 5.

Quantitative immunoblotting is challenging when imaging by chemiluminescence. (A to C) HT-29 lysates were prepared as in Fig. 4, immunoblotted for the indicated proteins, and imaged by IRDye fluorescence, enhanced chemiluminescence (ECL), or SuperSignal West Femto chemiluminescence as described (12, 16, 45, 46). Linear fits are shown in gray when the hyperbolic model is not significantly better than the linear model for that imaging condition. Linear fits are shown in red when the linear fit of the associated imaging condition is significantly better than the linear fit of the other imaging conditions. Hyperbolic fits are shown in green when the hyperbolic model is significantly better than the linear model for that imaging condition. Data are interpolated in blue when neither the linear nor the hyperbolic model provides a better fit. All model comparisons were done by the F test at a false-discovery rate of 5% (n = 4–8 dilutions). See file S2 for raw images and calcuations.