Figure 3.

Analysis of human gastric extracts after boiling with SDS/EDTA (M_C-406A) or after TRIzol^® extraction (M_C-577), respectively. (A) High-molecular-mass TFF2 was purified via SEC from the human gastric corpus, similar to as shown in Figure 1. Then, the corresponding fractions (B6–B10) were boiled with 1% SDS + 30 mM EDTA and separated immediately on a Superdex 75 HL column again. Elution profile after SEC as determined by absorbance at 280 nm (PAS-positive mucin fractions: pink). Underneath: Distribution of the relative TFF2 content. (B) Alternatively, a human gastric corpus specimen was extracted with TRIzol^®. Elution profile after SEC on a Superdex 75 HL column (PAS-positive fractions: pink). Underneath: Distribution of the relative TFF2 content. (C,D) Western blot analysis concerning TFF2 (15% SDS-PAGE), MUC6 (lectin GSA-II, 1% AgGE), and MUC5AC (1% AgGE), respectively, of the SDS/EDTA extract (C) or the TRIzol^® extract (D). Shown are also the hybridization signals (autoradiography) obtained after incubating the blots with ¹²⁵I-labeled recombinant human non-glycosylated TFF2 (C) or porcine pancreatic TFF2 (D) (overlay assays). The start of the AgGE is marked with a dot on the left.