To elucidate the nature of the CMS, we performed a biopsy of the right anconeus muscle at the age of five, which included in vitro microelectrode recordings and electron microscopy of the NMJ.
To elucidate whether the acquisition of CSC-like properties occurs through selection of pre-existing subclone(s) or stochastic adaptation of random cells in evolutionary response to microenvironmental CMS, we performed limiting dilution of parental cells in 96 wells and randomly chose 10 single cell-derived clones and repeated the CMS-simulating culture as described in Figure 3a.
To compare the standardized coral recruit density among atolls (densities of recruits standardized by the mean atoll-specific total densities of colonies <5 cm) we performed ANOVA on arcsine square root transformed data.
To identify genes with significant transcriptional changes associated with CM, we performed, on the whole data set, a multi-class SAM procedure with a very stringent false discovery rate followed by a Welch t test with a Bonferroni correction.
To understand the different behaviours in VL hiPSC-CM and hAdultV-CM, we performed additional in silico analyses to evaluate the role of I Kr in compensating for the lack of a repolarizing effect of I K1 in hAdultV-CM versus VL hiPSC-CM.
To determine whether the chemokines MCP-1, -2 and -3 produced by BM stromal cells are involved in the chemoattractive effect of BM CM, we performed migration experiments with MM cells towards BM CM in the presence of a blocking CCR2 MoAb or neutralising MCP MoAbs.
To rule out that the ECS used in the DRG experiments may interfere with CM actions, we performed transport assays with HeLa cells using the ECS as for the DRG experiments.
Briefly, after seeding HEK293T cells in a 10-cm dish, we performed a triple transient transfection with the respective transfer plasmids, a packaging plasmid and an envelope plasmid encoding VSV G.
In one patient with a cyst length in the bone axis of 14 cm (Fig. 3), we performed a two-stage monolocal compression distraction osteogenesis, first in the distal and then in the proximal part of the cyst, given the poor infilling of distal and proximal areas of the cystic cavity.
If the tumors were larger than 2.5 cm in greatest diameter, we performed multiple overlapping ablations to cover the entire tumor, with a 5 mm ablative margin.
After the anastomosis, if the anastomotic stoma was within 2 3 cm from the dentate line, we performed a protective stoma, but if the anastomotic stoma was more than 3 cm from the dentate line, we did not perform a protective stoma.
