Erythrocyte-mediated microinjection was used to introduce [35S]polypeptides translated in vitro into 3T3-L1 cells. Such [35S]polypeptides are not degraded after loading into erythrocytes and are stable for the first 2 h after microinjection into growing 3T3-L1 cells. Similarly, little or no degradation of microinjected [35S]polypeptides is observed in either growing or confluent 3T3-L1 cells ov...

The exceptional requirement of La Crosse virus mRNA synthesis for ongoing protein synthesis in vivo was examined in vitro by using purified virions and a reticulocyte lysate. Transcription from the S genome produced two incomplete transcripts (110 and 205 nucleotides [nt]) in the absence of the lysate, whereas S-mRNA (900 nt) was predominantly made when the lysate was present. The addition of d...

Research questionHow does platelet rich plasma products like human Platelet Lysate (hPL) and umbilical-cord blood (UCP) affect the growth survival of isolated pre-antral follicles in vitro?DesignHuman (n=724; mean diameter: 75 µm; range: 46-237 µm) were from ovarian medulla donated by 14 patients undergoing unilateral oophorectomy for tissue cryopreservation. Follicles encapsulated 0.5% alginat...

Background: Mesenchymal stromal cells (MSC) are being tested for the treatment of immune diseases. MSC are present in several adult tissues which milieu may influence MSC behavior particularly under inflammatory conditions. Additionally, culture conditions also can modify cell function or state of activation. Methods: To address the influence of the MSC source on its characteristics, we studied...

Interactions between DNA and transcription factors (TFs) guide cellular function and development, yet the complexities of gene regulation are still far from being understood. Such understanding is limited by a paucity of techniques with which to probe DNA-protein interactions. We have devised magnetic protein immobilization on enhancer DNA (MagPIE), a simple, rapid, multi-parametric assay using...

In the absence of hormone, the avian progesterone receptor exists in a large multiprotein complex that is inactive but able to bind and respond to progestins. This inactive complex can be reconstituted in vitro by incubation of receptor monomer in rabbit reticulocyte lysate in the presence of ATP and magnesium. This results in receptor binding to the two heat shock proteins, hsp90 and hsp70, th...

Lysate microarrays (reverse-phase protein arrays) hold great promise as a tool for systems-level investigations of signaling and multiplexed analyses of disease biomarkers. To date, however, widespread use of this technology has been limited by questions concerning data quality and the specificity of detection reagents. To address these concerns, we developed a strategy to identify high-quality...