Fig. 3.
Panel a: ELISA studies of HeLa cells show 2–3 times more phosphorylated PDGFRβ in cells transduced with the PDGFRB-Asn666Ser vector than with the PDGFRB- wild type (wt) vector. Panel b: Western blots of cultured fibroblasts from a healthy control (Ctr) and patient-1 (N666S) with the Asn666Ser variant, untreated or treated with 0.1 μM imatinib (STI57). Specific antibodies were used to detect phosphorylated PLCγ1 (P-PLCγ1) and total PLCγ1 (PLCγ1), phosphorylated STAT1 (P-STAT1) and total STAT1 (STAT), phosphorylated-Tyr542-PTPN11, (P-PTPN11Y542) phosphorylated-Tyr580- PTPN11 (P-PTPN11Y580, and total PTPN11 (PTPN11), phosphorylated AKT (P-AKT) and total AKT (AKT), phosphorylated MAPK3 and total MAPK3. The amount of GAPDH (lines marked with asterisk (*)) was measured as a protein loading control. Molecular sizes are indicated in kDa. Similar results were seen using skin fibroblasts from Patient-2