Dr. Vogelsang-Eastwood has found only one tear, in a section of a sash that would have been subject to pulling.
Lysates were subject to pull-down at 4 °C with 4X-ZFYVE-GST on Glutathione beads in order to enrich for activated RAB5.
While we cannot currently distinguish between these models, the behavior of the glands in sma-1 (and other) mutants suggests that gland projections are not subject to pulling forces, but rather push outwards, with the muscles normally functioning to constrain the projection and prevent overgrowth.
In the tests, one plain strip with smooth surface, two strips with simple ribs, and eighteen strips with transverse members of various depths and counts were subjected to pull out forces with normal stresses ranging from 50 kPa to 100 kPa applied.
Due to larger amount of forces transferred to column, end column stubs subjected to pull and push at top and bottom end, respectively.
To further confirm these observations we performed experiments in which total cell lysates from isoform 2 or isoform 3 expressing cell lines were subjected to pull down assays with GST tagged dominant negative (T17N) or constitutively active (Q61L) Rac1 mutants (Fig. 4A, right panel).
The cell extracts were subjected to pull down assays with GFP-Trap.
All ubiquitination reactions were stopped by denaturation with 1% SDS, diluted with Triton X-100 and subjected to pull down using immobilized Co2+.
(D ) Cell extracts from HEK293 overexpressed HA-TBC1D15 and YFP, YFP-GABARAPL1 (YFP-WT), or its Y49AL50A mutant (YFP-YL) were subjected to pull down assays with GFP-Trap.
(D ) Extracts from cells overexpressing both HA-TBC1D17 and YFP, YFP-GABARAPL1 (YFP-L1), its Y49AL50A mutant (YL), YFP-LC3B or its F52A/L53A mutant (FL) were subjected to pull down assays with GFP-Trap.
This paper presents an investigation into the failure mechanism and alternative design of composite sandwich T-joints subjected to pulling load.
