Figure 3

PLV-LMCs originate from a Prrx1+ and NG2+ progenitor cell similar to VSMCs prior to P21. Multicolor fluorescent microscopy was performed on whole mount immunostained PLVs and adjacent blood vessels (BVs) from constitutive Prrx1Cre (n = 3, P21 depicted) and NG2Cre (n = 4, P90 depicted) reporter mice as described in Fig. 1. Representative low-magnification overlay images of all channels show the PLVs with a highlighted region of interest (white boxes) (A.a,B.a). High-magnification images of the region of interest show Prox1 immunostain of LECs (A.b,B.b), αSMA immunostain of LMCs (A.c,B.c), Cre-driven tdT expression (A.d,B.d), overlay of all channels (A.e,B.e), and positive control BV VSMCs directly adjacent to PLVs (A.f,B.f). Note tdT expression (red) in PLVs (A.d,B.d) that colocalizes with αSMA+ LMCs (A.e,B.e), which is more profound (yellow) on the VSMC dense BVs, as expected (A.f,B.f).