Figure 2.
LOX deletion in male MKs impacts bone collagen structure but not level, with associated changes in MK gene expression. (A) Representative images of bone sections of Pf4Cre+/+ and Pf4Cre+/+/LOXfl/fl male mice stained with picrosirius red and visualized under polarized light. Picric acid can more easily dissolve into collagen fibers that are loosely packed, yielding a bright yellow birefringence, as observed under polarized light. When the stain is less able to penetrate collagen because of tight fibril packing, collagen profile appears less bright under polarized light, with birefringence mixed with evidence of green, yellow, and sometimes orange/red. Five Pf4Cre+/+/LOXfl/fl and 3 Pf4Cre+/+ 17-week-old male mice were analyzed. Images shown are representative of at least 3 slides analyzed in each case. (B) Representative images of bone sections of Pf4Cre+/+ and Pf4Cre+/+/LOXfl/fl male mice stained with picrosirius red and visualized under nonpolarized light for total collagen detection. Five Pf4Cre+/+/LOXfl/fl and 3 Pf4Cre+/+ 17-week-old male mice were analyzed. Images shown are representative of at least 3 slides analyzed in each case. (C) Differential expression of bone development–related genes in MKs of Pf4Cre+/+/LOXfl/fl mice compared with Pf4Cre+/+ mice with a graphical representation of >twofold difference in expression, using the commercial RT2 Profiler PCR Array Mouse Osteogenesis PAMM-026Z. mRNA results were normalized to β-actin, β-2 microglobulin, glyceraldehyde-3-phosphate dehydrogenase, β-glucuronidase, and heat shock protein 90-α. Eight 12-week-old control mice and 8 12-week-old Pf4Cre+/+/LOXfl/fl mice were analyzed. MKs derived from 4 mice in each experimental group were combined as 1 sample to obtain sufficient material and increase biological representation. Ahsg, alpha-2-HS-glycoprotein; Alp, alkaline phosphatase; Bmp, bone morphogenetic protein; COl, collagen; Csf, colony stimulating factor; Fgfr, fibroblast growth factor receptor; Fn, fibronectin; Mmp, matrix metallopeptidase; Pdgfa, platelet derived growth factor alpha; Tnf, tumor necrosis factor.

LOX deletion in male MKs impacts bone collagen structure but not level, with associated changes in MK gene expression. (A) Representative images of bone sections of Pf4Cre+/+ and Pf4Cre+/+/LOXfl/fl male mice stained with picrosirius red and visualized under polarized light. Picric acid can more easily dissolve into collagen fibers that are loosely packed, yielding a bright yellow birefringence, as observed under polarized light. When the stain is less able to penetrate collagen because of tight fibril packing, collagen profile appears less bright under polarized light, with birefringence mixed with evidence of green, yellow, and sometimes orange/red. Five Pf4Cre+/+/LOXfl/fl and 3 Pf4Cre+/+ 17-week-old male mice were analyzed. Images shown are representative of at least 3 slides analyzed in each case. (B) Representative images of bone sections of Pf4Cre+/+ and Pf4Cre+/+/LOXfl/fl male mice stained with picrosirius red and visualized under nonpolarized light for total collagen detection. Five Pf4Cre+/+/LOXfl/fl and 3 Pf4Cre+/+ 17-week-old male mice were analyzed. Images shown are representative of at least 3 slides analyzed in each case. (C) Differential expression of bone development–related genes in MKs of Pf4Cre+/+/LOXfl/fl mice compared with Pf4Cre+/+ mice with a graphical representation of >twofold difference in expression, using the commercial RT2 Profiler PCR Array Mouse Osteogenesis PAMM-026Z. mRNA results were normalized to β-actin, β-2 microglobulin, glyceraldehyde-3-phosphate dehydrogenase, β-glucuronidase, and heat shock protein 90-α. Eight 12-week-old control mice and 8 12-week-old Pf4Cre+/+/LOXfl/fl mice were analyzed. MKs derived from 4 mice in each experimental group were combined as 1 sample to obtain sufficient material and increase biological representation. Ahsg, alpha-2-HS-glycoprotein; Alp, alkaline phosphatase; Bmp, bone morphogenetic protein; COl, collagen; Csf, colony stimulating factor; Fgfr, fibroblast growth factor receptor; Fn, fibronectin; Mmp, matrix metallopeptidase; Pdgfa, platelet derived growth factor alpha; Tnf, tumor necrosis factor.

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