Figure 2.

Delivery of Ectopic Human FGFR2 Alleles in Adult Mouse SSCs

(A) Schematic of FGFR2 protein and primers for human-specific FGFR2 qRT-PCR. Asterisk marks the location of the S252W Apert mutation (ECD, extracellular domain).

(B) qRT-PCR using two sets of primers showing relative expression levels of hFGFR2 in lentivirus-transduced SSCs bearing WT hFGFR2 (WT) or human S252W FGFR2 (S252W) with untransduced SSCs as the reference control (mean ± SD of three independent cell lines per genotype).

(C) IF on cytospins (top row) with anti-hFGFR2 antibody (green) for SSCs stably expressing WT (middle) or S252W (right) alleles and untransduced SSCs as control (left). Scale bar, 10 μm. Phase contrast (bottom row) of untransduced SSCs (control; left) or stably expressing hFGFR2 WT (middle) or S252W (right) alleles in standard culture conditions (scale bar, 50 μm).

(D) Immunoprecipitation (IP) and detection with two specific monoclonal antibodies for hFGFR2 in WT and S252W SSC. Controls: untransduced SSCs and JK1 feeder cells or 293T cells transduced with WT or S252W lentiviruses. Grey dashed line denotes cropped lane.

See also Figure S1.