(A)
Left to right, IF staining of menin in duodenums of omeprazole (OM) treated C57 WT, Men1Δ
IEC mice, untreated Men1ΔIEC;Sst−/−, and OMS mice. Far right panel, 4 months after withdrawal (WD) of OM from OMS mice. (B) IF staining of menin and gastrin in glial cultures isolated from duodenal lamina propria of untreated Men1ΔIEC;Sst−/− and OMS mice. (C) Representative blots showing menin expression (top) in total cell lysates (β-actin loading control), nuclear (Sp1 loading control), and cytoplasmic fractions (GAPDH loading control) of glial cultures from mice; quantitation of menin expression (bottom) in nuclear and cytoplasmic fractions of glial cultures, expressed as integrated band intensities normalized to loading controls (n=7–9 mice). (D) IF staining of gastrin and CCKBR in glial cultures from untreated Men1ΔIEC;Sst−/− (−OM, top panel) and OMS mice (+OM, bottom panel) mice. (E) Representative blot (top) and quantitation of CCKBR expression (bottom) normalized to GAPDH in glial cultures isolated from mice (n=7–9 mice). (F) Representative blot showing menin expression in nuclear (Sp1 loading control) and cytoplasmic fractions (Gapdh loading control) of glial cultures from untreated C57 WT mice treated without or with 20 nM gastrin in the presence or absence of YM022. Data shown are the Median ± Interquartile Range of median. *** p< 0.001, **** p< 0.0001. Scale bars: (A) - 50 μm, (inset in A) – 20 μm, (B, E) – 20 μm.