A second morpholino (MO-2), designed upstream to MO-1, mimicked this phenotype, but the effect was milder (data not shown).
Interestingly, in contrast to prophase anaphase when the spindle vigorously oscillated vertically, during the process of centrosome attachment to the cortex, spindle oscillation became milder (data not shown) and posterior centrosomes were slightly elongated toward the P2/EMS boundary (Fig. 1b; 2:00).
In an initial characterization, we found that ahsa1 was elevated 3.4-fold in severe mutants relative to mild mutants (data not shown).
Surprisingly, we found that inhibition of Hsp90 caused the partial restoration of the trabeculae in the severe genetic background (Fig. 2) but did not alter the craniofacial phenotype of mild mutants (data not shown).
In MTT assays, ZOL at 10 μ M exhibited mild toxicity (data not shown); however, this toxicity was accounted for in CSAs, as surviving fraction is normalised to unirradiated control for each concentration of ZOL used.
In the newly diagnosed group, two out of the four urine samples were negative for PSA mRNA transcripts, before mild prostate massage, whereas all were positive after mild prostate massage (data not shown), indicating that mild prostate massage increased the exosomal secretion into the urethra and subsequently into the collected urine fraction.
However, all mutants showed mild sensitivity to an acute exposure to another superoxide generating agent, juglone and mild heat-shock (data not shown).
It should be noted that CD9 was not observed to associate with integrin α2β1 by immunoprecipitation experiments after cell lysis using the mild detergent Brij58 (data not shown).
Treatments performed at later times resulted in an almost normal development of Mc with only mild proximal defects (data not shown).
Although chelation studies have been helpful in defining a role for zinc in many biological processes, we have found that hESCs are highly sensitive and do not survive even very mild chelation conditions (data not shown).
