Figure 2 | Bone Research

Figure 2

From: The heterodimeric structure of heterogeneous nuclear ribonucleoprotein C1/C2 dictates 1,25-dihydroxyvitamin D-directed transcriptional events in osteoblasts

Figure 2

Stable expression of only human hnRNPC1 protein in mouse MC3T3-E1 osteoblasts does not confer resistance to 1,25(OH)2D. (a) Osteoblastic transduction was performed using the lentivirus RRL transfer vector harboring a stop codon-less human (hu) hnRNPC1 cDNA and a GFP cassette driven by the bone-specific mouse col1a1 2.3 kB promoter. The lentivector backbone also contains the cPPT from the HIV-1 integrase gene to increase the copy number of lentivirus integrating into the host genome to enhance viral titer. (b) FACS sorting of GFP-positive MC3T3-E1 osteoblasts transduced with the hu hnRNPC1 and empty lentivirus vectors for the isolation and generation of stable clones. (c) Fifteen hu hnRNPC1-overexpressing stable lines were created (data not shown), whereby five (clones 3″, 5″, 8″, 10″, 11″) lines with variable hnRNPC1 overexpression were further evaluated for 1,25(OH)2D (10 nmol·L−1, 6 h) mediated induction of endogenous mouse (mu) Cyp24a1 mRNA. cPPT, central polypurine tract.

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