Fig. 7.

Pharmacological blockade of Per1 nuclear entry leads to decreased occupancy of CLOCK and Per1 on the SGLT1 promoter in HK-2 cells. Chromatin immunoprecipitation experiments were performed using HK-2 cells treated with either vehicle (water) or 10 μM PF670462 for 24 h. End-point PCR was performed using primer sets flanking three putative E boxes in the SGLT1 promoter (A). B: input experiments. Chromatin immuprecipitations were performed using anti-Pol II (Santa Cruz Biotechnology; C), anti-CLOCK (Pierce; D), anti-Per1 (Pierce; E), or rabbit IgG (Bethyl, negative control; F) antibodies. Bands were quantitated using densitometry, which was performed using ImageJ (http://rsbweb.nih.gov/ij). Signal strength was normalized to the relevant vehicle- or CKinh-treated input control. Values are means ± SE; n = 3. *P < 0.05.