This new approach uses a two-stage model, which is able to capture all of the important experimentally observed effects, i.e. correct load transfer between film and substrate before and after cracking of the film, shear-stress-induced local out-of-plane deformation of the film and mixed mode delamination.
Phosphorylation of signalling proteins was corrected for loading anomalies to α-actin.
Subsequently samples were corrected for loading using the control band and finally values were expressed relative, defining the intensity of the wild type sample as 1.
Northern blots of total cellular RNA were quantified using the gel analysis function of ImageJ [50] and values for each protein corrected for loading errors by comparison with actin controls, and expressed as a fraction of paired control (DMSO) and experimental (10 µM hippuristanol) samples.
Western blots were quantified using the gel analysis function of ImageJ [50], and values for each protein corrected for loading errors by comparison with tubulin and actin controls, and expressed as a fraction of paired control (DMSO) and experimental (10 µM hippuristanol) samples.
The densitometric values are corrected for loading control shown in the bottom panel of the figures.
Protein measurements were corrected for loading as described [ 15- 17, 19].
Autoradiographs were scanned on a Umax Astra 1220 U scanner, quantified using ImageJ and corrected for loading using Erk1 [ 31].
Densitometry was performed using molecular analyst software (Bio-Rad, Hercules, CA), and all values were corrected for loading with actin.
Bands were quantitated by densitometry using the Molecular Dynamics Software ImageQuant GE Healthcare Life Sciencess, Piscataway, NJ, USA) and densitometric values were corrected for loading control.
