Research Paper Volume 12, Issue 24 pp 24853—24871

Aberrant RhoA activation in macrophages increases senescence-associated secretory phenotypes and ectopic calcification in muscular dystrophic mice

Figure 2. CD68+ cells at the sites of muscle calcification in dKO mice are positive for RhoA expression. (A) Immunostaining of dKO skeletal muscle (gastrocnemius) sections with RhoA and CD68 antibodies indicating that many of the accumulated CD68+ cells at sites of ectopic calcification (EC) are also RhoA+. (B) Immunostaining of dKO skeletal muscle and bone marrow sections with RhoA and CD68 antibodies showing that CD68+ cells distant from ectopic calcification or those found in bone marrow are negative for RhoA expression. (C) Immunostaining of dKO skeletal muscle with RhoA/F4-80 or RhoA/CD163 antibodies indicating RhoA+ cells are usually positive for F4-80, but negative for CD163 (a marker for M2 macrophages). (D) Quantification of RhoA+/CD68+ cells among CD68+ cells at locations of muscle with or without EC formation. * indicates p<0.05.