, endogenous CCK2R mRNA abundance in AR42J cells in response to serum withdrawal for times indicated

<p><b>Copyright information:</b></p><p>Taken from "Regulation of mammalian gastrin/CCK receptor (CCK2R) expression and "</p><p></p><p>Experimental Physiology 2007;93(2):223-236.</p><p>Published online 12 Oct 2007</p><p>PMCID:PMC2253704.</p><p>© The Authors. Journal compilation © 2007 The Physiological Society</p> Results are normalized to 18S rRNA and are expressed as percentage of full serum value (= 100%). = 4. * = 0.016, ** = 0.013, *** = 0.005 time 0, ANOVA. , CCK2R mRNA abundance in RGM1 cells in response to serum withdrawal for times indicated. Results are normalized to 18S rRNA and are expressed as percentage of full serum value (= 100%). = 3. * = 0.0016 time 0, ANOVA

, CCK2R immunoreactivity and DAPI staining in full-thickness corpus 2, 6, 9 and 13 days following cryoulcer generation

<p><b>Copyright information:</b></p><p>Taken from "Regulation of mammalian gastrin/CCK receptor (CCK2R) expression and "</p><p></p><p>Experimental Physiology 2007;93(2):223-236.</p><p>Published online 12 Oct 2007</p><p>PMCID:PMC2253704.</p><p>© The Authors. Journal compilation © 2007 The Physiological Society</p> Note intense epithelial staining at ulcer repair margins at 13 days, and staining in the submucosa beneath the ulcer crater (indicated by white line) at 9 days. Scale bar represents 500 μ. Sections are representative from four individual animals at each time point. , CCK2R and smooth muscle α-actin immunoreactivity in gastric corpus epithelium at the repair margin of a 9 day ulcer. Upper panel, CCK2R (FITC); centre panel, smooth muscle α-actin (Texas Red); and lower panel, overlay. , CCK2R and vimentin immunoreactivity in gastric corpus submucosa beneath the repairing crater of a 9 day ulcer. Upper panel, CCK2R (FITC); centre panel, vimentin (Texas Red); and lower panel, overlay. , deconvolved images of gastric corpus epithelial cells at the repair margin of a 9 day ulcer. Upper left panel, CCK2R (FITC); upper right panel, smooth muscle α-actin (Texas Red); and lower panel, overlay. , deconvolved images of gastric corpus submucosal cells beneath the repairing crater of a 9 day ulcer. Upper panel, CCK2R (FITC); centre panel, vimentin (Texas Red); and lower panel, overlay. For and , scale bar represents 50 μm; for and , scale bar respresents 10 μm

, CCK2R expression in mucosa and submucosa 9 days following cryoulcer generation

<p><b>Copyright information:</b></p><p>Taken from "Regulation of mammalian gastrin/CCK receptor (CCK2R) expression and "</p><p></p><p>Experimental Physiology 2007;93(2):223-236.</p><p>Published online 12 Oct 2007</p><p>PMCID:PMC2253704.</p><p>© The Authors. Journal compilation © 2007 The Physiological Society</p> Bracket indicates residual ulcer crater. , CCK2R expression in mucosa and submucosa 9 days following cryoulceration, 0.5 cm away from the ulcer site. C, regenerative mucosa, adjacent to cryoulcer. , submucosa below residual ulcer crater. , normal mucosa, 0.5 cm away from site of ulcer. , normal submucosa, 0.5 cm away from site of ulcer. For and , scale bar respresents 300 μm; for , scale bar represents 50 μm. Hybridizations were performed using a mixture (1:1:1) of the three oligonucleotides described in the Methods

, sequence of the human CCK2R promoter −130 to −190 region, showing consensus -regulatory elements

<p><b>Copyright information:</b></p><p>Taken from "Regulation of mammalian gastrin/CCK receptor (CCK2R) expression and "</p><p></p><p>Experimental Physiology 2007;93(2):223-236.</p><p>Published online 12 Oct 2007</p><p>PMCID:PMC2253704.</p><p>© The Authors. Journal compilation © 2007 The Physiological Society</p> Numbering is relative to the start of transcription (+1), which is taken as 193 bp upstream of the start of translation (see main text). , basal activity of CCK2R promoter–reporter constructs (size as indicated) 24 h after transfection in AGS cells. * Significantly reduced relative to 1700 bp construct ( < 0.001, = 3, ANOVA). , basal activity of CCK2R promoter–reporter constructs (size as indicated) 24 h after transfection in RGM1 cells. * Significantly reduced relative to 928 bp construct ( < 0.001, = 3, ANOVA). , basal activity of mutated 196 bp constructs 24 h after transfection into AGS cells. Mutations as indicated were made corresponding to the -regulatory elements underlined in . Values are percentage of wild-type (WT) sequence activity (= 100%). * Significantly reduced compared with WT activity ( < 0.001, = 3). , basal activity of mutated 196 bp constructs 24 h after transfection into RGM1 cells. Mutations as described in . Values are percentage of WT sequence activity (= 100%). * Significantly reduced compared with WT activity ( < 0.001, = 3, ANOVA)

Gastrin-responsiveness of the <i>PAI-2</i> and <i>Reg1</i> promoters following PSMA5 knockdown.

<p>Response of the <i>PAI-2</i> (left panel) and <i>Reg1</i> (right panel) promoters to gastrin (G17, 2×10⁻⁹ M) in AGS cells transfected with PSMA5 siRNA (KD) or scrambled RNA (Scr). Open bars, unstimulated cells; closed bars, cells stimulated with gastrin (G17, 2×10⁻⁹ M) for 6 h. Values are mean ± SEM, n = 4–6.</p

Knockdown of proteasome subunits in AGS-GR cells.

<p>Representative western blots of PSMB1 (left panel) and PSMA5 (right panel), in AGS-GR cells 72 hours after transfection with validated siRNA or scrambled control. Left lanes, scrambled control (Scr), right panels, knockdown (KD). Blots were re-probed for HSP90.</p

Nuclear and cytoplasmic abundance of proteasome subunits in gastrin-stimulated AGS-GR cells.

<p>A, representative western blot of proteasome subunits in cytoplasmic (left panel) and nuclear (right panel) extracts of AGS-GR cells, 0, 60, 90 or 120 minutes after stimulation with gastrin (G17, 2×10⁻⁹ M). Blots were re-probed with HSP90 (cytoplasmic) or lamin (nuclear). B, signals at 0 (control) and 120 (gastrin) minutes of gastrin stimulation were quantified by densitometry and normalized to HSP90 or lamin. Nuclear to cytoplasmic ratios at 0 min ( = 1.0) versus 120 min are shown for PSMB1, PSMA5 and PSMC1. ***, p<0.001; n = 7, ANOVA.</p

Gastrin-responsiveness of the <i>PAI-2</i> and <i>Reg1</i> promoters following PSMB1 knockdown.

<p>Response of the <i>PAI-2</i> (left panel) and <i>Reg1</i> (right panel) promoters to gastrin (G17, 2×10⁻⁹ M) in AGS cells transfected with PSMB1 siRNA (KD) or scrambled RNA (Scr). Open bars, unstimulated cells; closed bars, cells stimulated with gastrin (G17, 2×10⁻⁹ M) for 6 h. Values are mean ± SEM, n = 5–9. ***, p<0.001; *, p<0.05.</p