Fig. 6

Functional studies related to loss of BMPR2 in human monocytes. Induced pluripotent stem cells (iPSC) derived from HPAH patients or Controls were differentiated into induced monocytes (iMono) using a stepwise differentiation protocol outlined in the Methods. iMono of HPAH patients and Controls were compared for (A) CD14 cell surface expression, assessed by FACS; (B) Number of iMono per mL. on day 19 of differentiation. (C)STAT1 gene expression by RT-qPCR. (D) Schematic demonstrating iEC and iMono co-culture approach for assessing adhesion. iMono adhesion to Control or HPAH iEC, per field of view. (E) Schematic demonstrating iEC and iMono co-culture approach for assessing transendothelial migration. Migrated of iMono across Control or HPAH iEC. (F) VE-cadherin and ICAM1 expression in co-culture of iMono and iEC with quantification based on mean fluorescent intensity (MFI) of 10 FOV for each replicate. Representative images of the iEC/iMono co-culture including phase, DAPI (blue) VE-cadherin (CDH5) (green) and ICAM1 (red). Red dotted outline on phase and DAPI images show adhered iMono. White dotted outline on merged images show corresponding iMono (pink). Bars represent mean ± SEM. For all experiments, n = 3 biological replications. + denotes the minimum achievable P-value for n = 3 was reached by the non-parametric t test