Extended Data Figure 5. Effects of targeted miR-34a deletion.
a-d, Targeted miR-34a/b/c triple KO (34abc-TKO) mice were compared with WT control mice (5 month males, n=4). a-c, Bone marrow osteoclast differentiation assay. a, Expression of miR-34a was diminished while expression of miR-34b and miR-34c remained absent/low in osteoclast precursors on d3. b, Expression of osteoclast markers were increased. c, Number, size and resorptive activity of mature osteoclasts were increased. d, Serum CTX-1 was increased. e-h, Targeted full miR-34a KO (34a-full-KO) mice were compared with WT control mice (2 month females, n=3). e-g, Bone marrow osteoclast differentiation assay. e, Expression of miR-34a was diminished in osteoclast precursors on d3. f, Expression of osteoclast markers were increased. g, Number, size and resorptive activity of mature osteoclasts were increased. h, Serum CTX-1 was increased. i-n, Conditional miR-34a KO mice by Tie2-cre (34a-Tie2-KO) were compared with littermate miR-34af/f control mice (2 month males, n=6). i-k, Bone marrow osteoclast differentiation assay. i, miR-34a expression was reduced in osteoclast precursors on d3. j, Expression of osteoclast markers were increased. k, Number, size and resorptive activity of mature osteoclasts were increased. l, Serum CTX-1 was increased. m, Trabecular BV/TV of proximal tibiae by μCT. n, Histomorphometry of the distal femur and vertebrae. For c, g, k, Mature osteoclasts were identified as multinucleated (>3 nuclei) TRAP+ (purple) cells. Scale bar, 25μm. Quantification of osteoclast number/well is shown in black. Quantification of osteoclast resorptive activity by calcium release from bone to culture medium (μM) is shown in blue.