Possible Linkage of SP6 Transcriptional Activity with Amelogenesis by Protein Stabilization
Figure 1
Downregulation of SP6 in C9 cells during long-term culture. (a) SP6 expression in C9 cells (P7 and P28) and in parent G5 cells. SP6 protein levels were analysed by WB using anti-HA antibody. Arrow: SP6; asterisk: nonspecific band and Posi: positive control. (b) Sp6 mRNA expression in C9 cells (P7 and P28) and G5 cells. Upper panel indicates schematic structure of the Sp6 gene and expression plasmid together with primer positions. Primers designed to detect three distinct types of Sp6 mRNA. Arrows indicate each primer’s position and direction. Grey box: untranslated region; striped box: HA-tag; cds: coding region of Sp6; endo: endogenous Sp6; exo: exogenous Sp6. Middle panel shows the representative result of RT-PCR analysis. Lower panel shows densitometric analysis data of RT-PCR. Each Sp6 level was normalized by 18S rRNA level, and the relative ratio was calculated compared to that of G5 cells. The level of G5 cells was assigned as one.
