Osteoclast formation formed by CD11b+ cells from WT, p62KI, TIL-6, p62KI/TIL-6, and MVNP mice. (A) OCL formation by 1,25-(OH)2D3. Data are expressed as the mean ± SD (n = 4); *p < 0.01, significantly different from OCLs formed with the same treatment in WT mouse cultures. (B) OCL formation by RANKL. Data are expressed as the mean ± SD (n = 4); *p < 0.01, significantly different from OCLs formed with the same treatment in WT mouse cultures. (C) Nuclei per OCL. The nuclear number per OCL was determined by randomly scoring 25 OCLs formed in 1 × 10−8 M 1,25-(OH)2D3 or 100 ng/mL of RANKL-treated cultures. Data are expressed as the mean ± SD (n = 25); *p < 0.01, significantly different from OCLs formed with the same treatment in WT mouse cultures. (D) TAF12 expression in OCLs. CD11b+ mononuclear cells were treated with 10 ng/mL of M-CSF for 3 days, then cultured with RANKL (100 ng/mL) for 4 days and cell lysates were collected. TAF12 expression was analyzed by immunoblot using antibodies recognizing TAF12 (ProteinTech). TFIIB was used as a loading control. WT = wild-type; RANKL = receptor activator of NF-κB ligand; OCL = osteoclast; 1,25-(OH)2D3 = 1,25-dihydroxyvitamin D3; M-CSF = macrophage colony-stimulating factor; TFIIB = transcription factor IIB.