Fig. 1.
a Targeting strategy used to generate the Manf−/− (‘knockout first’) and Manffl/fl Col2Cre+ (conditional knockout allele) lines. Black boxes represent Manf exons; LoxP sites are marked by arrowheads and FRT sites by ovals. b Western blotting showing the deletion of MANF in Manf−/− liver at E19.5, GAPDH as loading control. c LacZ staining of the whole mount ß-galactosidase (Rosa26) reporter crossed with Col2Cre line embryo at E15.5, showing cartilage-specific expression of the Col2Cre transgene. d Western blotting showing deletion of MANF in Manffl/fl Col2Cre+ cartilage at P21, GAPDH as loading control. Scale bars 1 mm and 100 μm