Maintenance of S2 cells, construction of expression plasmids, and cell-adhesion/spindle orientation assays were performed as detailed previously (Johnston et al., 2011).
Similarly, the majority of lines that showed chaining behavior did not have a significant change in sexual orientation in the mate preference assay but, rather, appeared to have an overall decrease in the sex specificity of their courtship.
"The authors describe a 'strong correlation' between the tubulin staining (presented) which defines MT orientation, and EB1 assay data (not presented), which can show both MT orientation and polarity.
To achieve specificity, during upright orientation of the assay tubes, the buoyant weight of the cubes applies a net downward force of 1.8 pN, as estimated by simple force balance, enough to break weak nonspecific interactions which are on the order of 0.1 1.0 pN.
(C F ) Evolution of spindle orientation function as assayed in cultured S2 cells that do not express endogenous Dlg protein.
Nevertheless, such experiments supported the formation of dimers rather than higher order species, and pairs showing this behavior (e.g., SYNZIP14 SYNZIP21, Supplementary Data 1) were further characterized using orientation and affinity assays (Table 1).
In order to detect the fraction of clones carrying the Mitf target DNA fragment inserted into pBluescript II KS in correct orientation, a colony PCR assay was performed with colonies of both strains.
miR-124 mutant males exhibited a normal repertoire of behaviors when paired with sexually mature Canton S (CS) female virgins in a standard courtship assay (including orientation toward the female, courtship song, tapping, licking, abdomen curling, and attempted copulation).
While FP blocks escape of paused Pol II, elongating Pol II can still clear the gene, and both changes can easily be followed by the sensitive and transcription orientation specific GRO-seq assay.
We identified G1 progeny in which the RMCE event had occurred by the loss of the y+ marker of the MiMIC gene trap cassette and determined which RMCE events were in the proper orientation using a PCR assay (Venken et al., 2011a).
