The slides were stained using Romanowsky staining (May-Grünwald - Giemsa) in an automated stainer (Sysmex XE-2100, Sysmex Corporation, Japan) to ensure constant staining quality.
Although Myosin 1E and β3-integrin were aligned in spots, their intensities did not vary in unison, especially during the contractile phase of spreading (P2) (Fardin et al., 2010) (supplementary material Movie 3; Fig. 2C white dotted arrow), when, Myosin 1E intensity gradually deceased, but a constant staining of β3-integrin was observed in patches.
The cyclic deformation tests were conducted under constant plastic stain amplitude mode and the evolution of the cyclic stress and cyclic hardening rate with cumulative plastic strain were monitored.
In contrast, the anti human RAMP1 844 antibody coupled to Alexa 488 showed clear and constant neuronal staining throughout the whole ganglia (Figure 2B); some neurons exhibited very intense fluorescence, while others were less intensely stained; both were evaluated as immunopositive.
The constant positive staining of smooth muscle cells of tumor vessels served as the positive internal control as previously described [ 26].
Interestingly, the pattern of Armadillo staining did not change during infection in flies depleted of EGFR in enterocytes, but instead a weak and constant Armadillo staining was observed in enterocytes.
Figures 2A and 2B show the immunostaining of a positive typical hyperplasia and a positive fibroadenoma, respectively: a minority of the cells show nuclear immunoreactivity that appeared associated with a constant, cytoplasmic staining.
Rabbit IgG (Cell Signaling Technology [#3900S]) was used at the same concentration as D8H2 to determine any nonspecific (ie, antibody constant region) staining inherent in the detection reagents or tissues or arising in tissues; rabbit IgG does not control for the unique GR antigen binding region of clone D8H2.
In fact, the expression was higher and concerning the vast majority of the cells, and exclusively nuclear in the cancer tissues, whereas breast fibroadenomas and typical hyperplasias showed a weaker expression, not present in the majority of the cells, with a constant cytoplasmic staining.
Constant exposure can stain teeth so use sparingly.
