96 Chapter 4 Occurrence of MET ectodomain shedding and regulated intramembrane proteolysis in oral squamous cell carcinoma cell lines and tissues Measured MET mRNA expression levels for the included controls (HeLa, PC3, HT-29, and LNCaP) were as expected (Fig. 3a). Moreover, all oral squamous cell carcinoma cell lines reliably express MET with varying levels (Fig. 3a). With the exception of SCC-25, immunoreactivities observed for D1C2 and A2H2-3 are in line with MET mRNA expression levels (Fig. 3a, b). Observed C-terminal p145β immunoreactivity under reducing conditions is similar to N-terminal MET p145β immunoreactivity under nonreducing conditions. Fig. 3: Occurrence of MET ectodomain shedding and regulated intramembrane proteolysis in oral squamous cell carcinoma cell lines. a Quantitative real-time PCR (qRT-PCR) results showing average MET fluorescence standardized to average HPRT1 fluorescence. b Immunoreactivities observed for p145N-term. MET (A2H2-3) under nonreducing conditions, immunoreactivities observed for p145C-term. MET (D1C2) under reducing conditions. c Specific immunoreactivities observed using D1C2 showing C-terminal MET protein products and fragments under reducing conditions that are the result of shedding and/or regulated intramembrane proteolysis (18, Supplementary Table 5). d Immunoreactivities for MET ectodomain shedding observed with A2H2-3. e Legend for observed mRNA expression levels, immunoreactivities using western blotting and immunoreactivities using ELISA. *Reference results retrieved from (18) for HeLa, PC3, LNCaP, and HT-29.
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