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Gene Ontology and GO Annotations
Description Files Reviews Ask a question Bundle discount. Antigen Information Accession Number: Q Gene ID: Target Species: Human. B p65 in whole lysates and nuclear extracts. Number of Targets Detected: 1. Species Detected: Human. Design Principle: Sandwich-based. Method of Detection: Colorimetric. Solid Support: well Microplate. Prepare all reagents and samples as instructed in the manual.
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IKBKB elisa kit. CHUK elisa kit. MAP3K8 elisa kit. HDAC1 elisa kit. BCL3 elisa kit. NR3C1 elisa kit. EP elisa kit. Acute Myeloid Leukemia Pathway antibodies.
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The procedure for preparation of cleared nuclear lysates was essentially that of Schreiber et al. Santa Cruz, CA. Blots were probed with affinity-purified antibodies. SA and p50 cat. SA were purchased from Biomol. Viewed by phase contrast microscopy the day after plating Fig. However, when these cells are examined again after subculture, they have a typical fibroblast spindle shape, and their cytoplasm has become highly refractile Fig.
These changes are reflective of the reorganization of the actin cytoskeleton. The differences between freshly isolated stromal cells and their subcultures with respect to competence for collagenase synthesis might be due to their transformation in culture, similar to the transformation they undergo in the corneal wound.
However, because primary stromal cells undergo many replications before subculture, it was also possible that the change was due to selection for a subcomponent of the cell population with a growth advantage under the culture conditions.
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To distinguish between these possibilities, freshly isolated cells were plated in replicate culture wells and their phenotype was monitored daily to determine whether transformation could occur without subculturing. Results of a representative experiment are depicted in Figure 2 and summarized in Table 1. However, stress fibers had begun to develop in cells viewed 2 days after plating Fig. The cells had also become spindle-shaped by day 3, appearing identical with subcultured cells cf. No further changes in the actin cytoskeleton were observed on day 4 data not shown.
Tale of two transcription factors: NF-кB and HIF crosstalk
Progress of these cells toward competence for collagenase expression in response to CB was scored by using immunofluorescent localization to identify individual cells synthesizing collagenase and then by quantifying the percentage of these cells in the total population Table 1. Because intracellular collagenase concentrates within secretory cell compartments, collagenase-positive cells could be identified easily by their brightly fluorescent perinuclear spots Fig. Besides the change in cell shape, two additional changes were correlated with the burst of competence acquisition.
The first was in cell replication: the number of cells in a culture well doubled daily through day 3, and then replication ceased as cells became confluent. The second involved a change in response to CB: day 1 and 2 cultures did not collapse in response to CB Figs.
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