Figure 2: Liver Bmp6 mRNA is increased in endothelial Bmp2 KO mice, but Bmp2 mRNA and protein are not increased in endothelial Bmp6 KO mice.

(A-B) Eight-week-old littermate female (black closed symbols) and male (black open symbols) control (Bmp2^wt/wt;Bmp6^wt/wt;Tek-Cre+), single endothelial Bmp2 KO (Bmp2^fl/fl;Bmp6^wt/wt;Tek-Cre+), single endothelial Bmp6 KO (Bmp2^wt/wt;Bmp6^fl/fl;Tek-Cre+), and double endothelial Bmp2 and Bmp6 KO (Bmp2^fl/fl;Bmp6^fl/fl;Tek-Cre+) mice were analyzed for (A) liver Bmp6 and (B, left) Bmp2 relative to Rpl19 mRNA by qRT-PCR. The average of the control mice was set to 1. (B, right) Serum BMP2 protein levels were quantitated by ELISA. (C-D) Four-week-old female (black closed symbols) and male (black open symbols) global Bmp6^−/− and littermate WT mice (Bmp6^wt/wt) were fed a matched, purified iron sufficient (Control, 48 ppm iron) or high iron (High Fe, 2% carbonyl iron) diet for 3 weeks, followed by analysis of (C) liver iron, (D, left) liver Bmp2 relative to Rpl19 mRNA by qRT-PCR, and (D, right) serum BMP2 protein levels by ELISA. For qRT-PCR analysis, the average of the control Bmp6^wt/wt mice was set to 1. (E-F) Four- to five-week-old WT C57BL/6 male and female mice from four independent cohorts (grey symbols: ○ male Hfe^wt/wt littermates; ◻◼ female and male Bmp6^wt/wt littermates; ▲ Jackson female WT; ◇ Taconic male WT) were fed a matched, purified low iron (Low Fe, 2–6 ppm iron), iron sufficient (Control, 48 ppm iron), or high iron (High Fe, 2% carbonyl iron) diet for 3 weeks, followed by analysis of (E) liver Bmp6 and (F, left) Bmp2 relative to Rpl19 mRNA by qRT-PCR. The average of the control diet-fed mice was set to 1. (F, right) Serum BMP2 protein levels were quantified by ELISA. For all graphs, individual data points are shown and bars represent mean ± SEM. Results were analyzed by one-way ANOVA with Tukey’s post-hoc test. Means without a common superscript differ significantly (P < 0.05).