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. 2021 Jan 15;30(1):72–77. doi: 10.1093/hmg/ddab014

Figure 3.

Figure 3

Kinase phosphorylation measured by Human Phospho-Kinase Array. Two different OPDKD and control fibroblasts, incubated for 6 h at 32°C, were compared to cells kept at 37°C. We defined a significant difference to be of >50% increased phosphorylation measured by chemiluminescence. Top left: p.(Asn666Tyr) fibroblasts compared to control at 37°C showed increased levels of GSK-3α/β, AKT 1/2/3 and WNK1. Top right: At 32°C, PRAS40, PDGFRB, STAT5 α/β and PLCγ1 were also significantly different. PDGFRB, STAT5α/β had weak bands, whereas the band representing PLCγ was barely visible. Bottom: When fibroblasts harvested at 32°C were compared to cells harvested at 37°C, increased levels were found for GSK-3α/β (with 1.73/2.81 increase), AKT 1/2/3 (3.16/6.15), PRAS40 (4.32/4.49) and WNK1 (2.54/1.98) in both p.(Asn666Tyr) cell lines. No differences were found in either of the control fibroblasts. We found that in one of the p.(Asn666Tyr) cell lines there were more proteins with increased activation. This, and its control fibroblast cell line, were early passage (P3) cells with a rapid growth pattern, in contrast to the other p.(Asn666Tyr) cell line that was P5 and growing slightly slower. P-STAT1 is not part of the Human Phospho-Kinase Array.