Using a low-resolution filtered starting model, we then carried out projection matching and angular reconstitution cycles (see 'Materials and methods'; Figure 5 figure supplement 1C, Table 4) and then refined a 24 Å TBC-DEG:αβ-tubulin map.
The lyophilization-reconstitution cycle caused approximately 10% soluble protein loss (data not shown); however, the efficacy of reconstituted BsAb was slightly higher than that of the original sample (Additional file 1: Figure S2).
At the beginning of the following reproductive season, the spent gonad undergoes a period of reconstitution, and the cycle starts again (Gosling 2003).
The digest of OS membrane protein (25 µg) in 200 µL of 0.5% PFOA in 100 mM ABC (total PFOA amount = 1 mg) was subjected to speed-vac, and then three cycles of reconstitution in 100 µL of ethanol∶ethylacetate∶water∶TFA (0.33∶0.33∶0.33∶0.01, v/v/v/v) and evaporation in a speed-vac concentrator under low pressure of <10 mTorr.
In order to remove the residual PFOA completely, the rest of the digest (22.5 µg) was subjected to another three cycles of reconstitution in 100 µL of ethanol∶ethylacetate∶water∶TFA (0.33∶0.33∶0.33∶0.01, v/v/v/v) and evaporation at 60°C under atmospheric pressure of 760 Torr.
We found that the PFOA amount in the BSA digest was decreased below detectable level after three cycles of reconstitution in 100 µL of ethanol∶ethylacetate∶water∶TFA (0.33∶0.33∶0.33∶0.01, v/v/v/v) and evaporation at 60°C under atmospheric pressure of 760 Torr (data not shown).
Cells exposed to high concentrations (5.0 mM) showed a different effect, the C- metaphase presented fibrous and branched cortical filaments that allowed reconstitution of 4C nuclei and cell cycle progression.
Interestingly, the mathematical model showed that recruitment of quiescent cells into the active cell cycle promotes β-cell mass reconstitution in the Cdk4R24C pancreas.
Understanding the specific conditions of use for the product life cycle, including production, distribution, storage and reconstitution will allow designers to choose materials that facilitate design.
Particle orientations were refined by multiple cycles of MRA, MSA and angular reconstitution, gradually incorporating more particles.
Particle orientations were refined in multiple cycles of MRA, MSA and angular reconstitution and the resulting 3-D reconstruction was used as an initial model for projection matching in SPIDER.
