Fig 1: Investigation of the XYLT mRNA and XT-I protein expression level in acute senescent proto-myofibroblasts. After 24 h of cultivation, cells were treated for 1 h with 300 µM H2O2 or H2O (control). XYLT1 (A) and XYLT2 (B) mRNA expression levels were analyzed after 6 or 72 h of cultivation by quantitative real-time PCR and were normalized on GAPDH mRNA expression level. The XT-I protein expressions in cell lysates were detected via immunoblotting 72 h after H2O2 treatment and were normalized on GAPDH protein expression. Quantification was determined using ImageJ (C). The intra- and extracellular XT-I enzyme activities were determined after 72 + 18 h by a XT-I-selective mass spectrometric assay (UPLC/ESI-MS/MS assay) and were normalized on total protein concentrations (D). The values shown are means ± SEM for three biological and three (A,B,D) or one (C) technical replicate per experiment (n = 3) and calculated relative to the respective control. Mann-Whitney U test: not significant (ns), p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).
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