Tool 10000 days rar11/9/2022 The RARa and PML-RAK immunoreactive bands were visualized with a ehemiluminescent substrate following incubation with a secondary antibody linked to horseradish peroxidase. (b) Protein homogenates (100 &lane) from NB4 cells culture8fo 'k days in medium alone or in medium containing 1,000 units/ml IFNa were electrophoresed on an 8% SDS-denatured polyacrylamide gel and subjected to Western-blot analysis with a polyelonal antibody recogniring the F domain of RARa. The white, stippled and black bars indicate the level of ex ression of RARal, RARa2 and PML-RAR transcripts respectively. All the data are normalized for the level of expression of the G3PD mRNA observed at each experimental point. Results are expressed in fold induction relative to the value determined for NB4 or HL-60 cells incubated in medium alone and taken as 1. The densitometric analysis of each RAKa or PML-RAR signal is indicated at the top of the autoradiograph. The position of the 2 RARa, PML-RAR and G3PD transcripts is indicated o n the left, that of the 28s and 18s rRNA on the right. Filters were sequentially hybridized with a cDNA coding for RARa and glucose-3-phosphate dehydrogenase (G3PD). (a) Total RNA was extracted from NB4 or HL-60 cells treated for 4 days in the ahsence o r in the presence of 100, 1,000 and 10,000 units/ml of IFNa or 1,000 units/ml of IFNp and used for Northern-blot analysis ( I 0 Fgieach lane). Effects of IFNa on RARa and PML-RAR mRNA and proteins in NB4 and HL-60 cells.
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