Vehicle-treated cells

Inhibition rate (%) = (A-B)/(Axl00), where A is the mean OD of vehicle treated cells and B is the mean OD of compound treated cells. Values are the mean standard deviation. (N.A., not active.) p <0.05, p <0.01 and /> <0.001 compared to control as determined by Tukey s Studentized Range (HSD) test. 

Seed 2x10 inhibitor- or vehicle-treated cells in 100 pL per well in 96-well plates. Incubate for 48 h at 37 °C and 5 % CO2. 

Count cells and make aliquots of 5 x 10 cells per 1.5 mL sterile microtube. Prepare six tubes for each target gene two tubes for the control non-targeting siRNA, and two tubes for each of the two specific siRNAs (the first tube is for vehicle treated cells the second tube is for PTP inhibitor treated cells). 

A single cell suspension of LNC is prepared under aseptic conditions and cultured for various periods of time. The production by LNC of type lcytokines (such as IFN-y and IL-12) and type 2 cytokines (such as IL-4, IL-5, IL-10 and IL-13) is measured using cytokine specific ELIS As or cytokine microarrays. It is possible to measure the spontaneous production of IL-5, IL-10, IL-12, IL-13 and IFN-y by LNC without restimulation in vitro. However, IL-4 appears to be produced in comparatively small amounts and to induce detectable levels of this cytokine an additional stimulus is required concanavalin A (con A), a T lymphocyte mitogen. Culture of LNC derived from mice treated with chemical respiratory sensitizer with con A will stimulate the production of detectable levels of IL-4. Treatment with the same mitogen of LNC derived from naive or vehicle-treated mice fails to induce measurable IL-4 secretion [86],... 

After behavioral evaluation in the 8-arm radial maze task, the rat s brain was fixed and apoptosis was identified in the hippocampal CA1 by TUNEL assay. Apoptotic cells were found in vehicle-treated rats on the 7th day after ischemia (mean F S.E.M. 78.4 F 5.7 TUNEL-positive cells/mm2). As shown in Fig. 8,... 

Figure 45- 5 Graphic example of glomerular changes in nephrotic syndrome. Scanning electron microscopic view of glomerular epithelial podocytes from a vehicle-treated rat (left) and a puromycin aminonudeoside (PAN)-treated ( 80mg/kg body wt) rat (right). Note the extensive loss of podocyte foot processes, which occurs in response to PAN-induced nephrotic syndrome. This illustrates the major cellular changes that can occur in nephrotic syndrome. GEC, glomerular epithelial cell. (From Ricardo SD, Bertrom JF, Ryan GB Antioxidants protect podocyte foot processes in puromycin aminonucleoside-treated rats. J Am Soc Nephrol 1994 4 1974-86.)...

Fig. 3.12. A Protective effect of (-)-BPAP in the high micromolar concentration range, with a peak effect at 10 6M concentration, against serum-free condition induced cell death in low-cell-density culture of the cerebral cortex from E17 rats. B Lack of a protective effect of (-)-BPAP under the same conditions in the low nanomolar concentration range. Experiments were carried out in triplicate. Data are the mean SEM from six independent experiments. The data were analyzed using Student s f-test after multiple comparisons of ANOVA. P value was < 0.05 compared with the results in the vehicle-treated culture. See Hamabe et al. (2000) for methodology...

Add 10 pg of purified anti-CD3 antibody to four tubes of each the vehicle- and inhibitor-treated cells and mix by pipetting up and down (one tube from each condition will be left... 

Chan and Delucchi, 2000 Desai et al., 2002 Goodwin et ah, 1999 He et al., 1999 Ledirac et al., 2000 Luo et al., 2002, 2003, 2004 Pichard et al., 1999 Zhao et ah, 2002). As a result, overall CYP3A4 activity in cells treated with these compounds can actually be lower than activity obtained from vehicle control cells, even though CYP3A4 mRNA levels and protein expression are significantly induced. 

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