Fig. 3: RACK1 regulates RANKL-induced c-Src activation.
From: RACK1 interaction with c-Src is essential for osteoclast function
a Mature osteoclasts generated from BMMs were serum-starved and stimulated with 200 ng/mL RANKL for 20 min. Whole cell lysates were analyzed using western blotting with anti-phospho-c-Src, anti-c-Src, anti-HA, and anti-actin. The ratio of p-c-Src to total c-Src proteins was quantified from each of three independent experiments. *P < 0.01. b BMMs transduced with pMX-puro control shRNA (control) or pMX-puro-shRACK1 (shRACK1) retrovirus were cultured for 3 days with 30 ng/mL M-CSF and 100 ng/mL RANKL to generate mature osteoclasts. Protein levels were analyzed using western blotting with antibodies specific for the indicated proteins. The ratios of p-c-Src to actin and RACK1 to actin were quantified from each of three independent experiments. *P < 0.01. c Mature osteoclasts generated from BMMs were cultured with 30 ng/mL M-CSF and 100 ng/mL RANKL for 4 days. After the cells were removed, they were either maintained in suspension or plated on a vitronectin-coated dish for 30 min. Cell lysates were analyzed using western blotting with antibodies against anti-phospho-c-Src, anti-c-Src, anti-HA, and anti-actin. The ratio of p-c-Src to total c-Src proteins was quantified in each of three independent experiments. *P < 0.01. Western blots in a–c are representative of three independent experiments
