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The reported stably CaSR-transfected HEK293 cell line (HEK-CaSR) (9, 11) was cultured in high-glucose DMEM (Invitrogen) supplemented with 10% fetal bovine serum and 1% geneticin (11).
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This promoter has previously been reported to stably express eGFP in both undifferentiated cells and derivatives of all three embryonic germ layers when inserted into to the Envy locus by transfection of bacterial plasmids[6].
A similar effect of CK overabundance in flowering delay was reported in stably transformed tobacco plants with 35S:: MdIPT3a [ 31].
We have previously reported that stably transfected RB functions as a growth and tumour suppressor in HTB9 human bladder carcinoma cells, which carry a mutation of the p53 gene at codon 280 and lack RB expression.
Recently 5'-half-tRNA fragments have been reported to exist stably in various types of cells, although little is know about their physiological roles.
ACT2/7 has been reported as a stably expressed gene whereas ACT11 was reported as unstable [ 38, 39].
In addition, PPIB was included because it was previously reported to be stably expressed in blood [ 3].
PPIB was also reported to be stably expressed in whole blood of patients with local and systemic inflammatory syndromes and healthy controls [ 3].
We have previously reported on the stably transduced PKA, Epac 1 and Epac 2 knockdown Raw 264.7 cells and shown that there was a marked reduction in the expression of these proteins [ 19].
Recently, Kim et al. reported that AuNPs stably functionalized with covalently attached oligonucleotides activate immune-related genes and pathways in human peripheral blood mononuclear cells, but not an immortalized, lineage-restricted cell line [ 87].
qRT-PCR validation was then performed on 10 candidate reference genes, four commonly used reference genes (ACTB, GAPDH, B2M, and HMBS) and PPIB, which is reported to be stably expressed in blood [ 3].
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