Figure 1.

Study design and representative gating strategy for isolation of pericyte types using fluorescence-activated cell sorting (FACS). A: pericytes were isolated from age-matched, mobile, C57BL/6J male donor mice 9–10 days before transplantation. Fourteen days after unilateral hindlimb immobilization, the staple was removed and either PBS or pericytes (NG2⁺Lin⁻, CD146⁺NG2-Lin⁻ or CD146⁺Lin⁻) were directly injected into the immobilized tibialis anterior (TA) muscle. The contralateral control served as the uninjected mobile control. Muscles were dissected and evaluated 14 days after remobilization, and a paired difference of zero indicates full recovery compared with the mobile control. B: the mononuclear cell population was identified based on forward scatter (FSC) and side scatter (SSC) size. After exclusion of debris and cell doublets, NG2⁺, CD146⁺NG2⁻, and CD146⁺ pericytes were gated from the lineage negative single cells (CD31⁻CD45⁻). Cells were recovered in culture for up to 10 days without splitting before transplantation. IM, immobilization; Lin, lineage; NG2, neural/glial antigen 2; PE, phycoerythrin; RE, remobilization.