Illumina HiSeq 2000 paired end sequencing; RNA-Seq of osteoclasts derived from female Phlpp1 cKO or control littermates
RNA-Seq of osteoclasts derived from female Phlpp1 cKO or control littermates
Samples were placed on ice, rinsed with PBS and lysed in Trizol. Hind limbs were dissected and bone marrow macrophages were collected from female 6 to 8-week-old Control, Phlpp1 cKOCtsk or Phlpp1-/-  mice as previously described (16). Briefly, cells were flushed, pelleted and red blood cells were lysed (RBC Lysis Buffer, #00-4333-57, Invitrogen, Carlsbad, CA). Cells were pelleted and cultured overnight in phenol red-free alpha MEM, 10% FBS and 1% antibiotic/antimycotic supplemented with 35 ng/mL M-CSF (#416-ML, R&D, Minneapolis, MN).  Non-adherent cells were then placed in culture medium supplemented with 35 ng/mL M-CSF, and 90 ng/mL RANKL (#315-11, Prepro Tech, Rocky Hill, NJ).  Cultures were fed on day 3 with culture medium plus 35 ng/ml M-CSF, and 90 ng/mL RANKL Trizol extraction Random hexamers were used to generate cDNA libraries via reverse transcription
UNIVERSITY OF MINNESOTA
E-MTAB-10487:WT_p