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NF-κB activation in Rab11a ΔIEC mouse intestines was also supported by elevated nuclear P65 levels from fractionated tissue lysates (Fig 4D).
While it is widely accepted that Wnt signaling influences proliferation in all crypts, the number of Wnt-activiated cells detected in Wnt-reporter mouse intestines was lower than expected.
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To determine if Wnt-receiving crypt cells might share expression with stem or early progenitor cells, Wnt-reporter mouse intestines were stained with antibodies for a putative intestinal epithelial stem cell marker, Musashi-1 (Msi-1) [ 35- 37].
Mouse intestines were fixed in 5% neutral-buffered formalin and embedded in paraffin.
Both mouse reporter lines displayed a similar pattern, therefore TOPGAL mouse intestines are depicted unless otherwise noted.
Mouse intestines were labelled by i.p. injection of 250 µl BrdU 2 and 48 h prior to tissue dissection.
Mouse intestines were dissected, washed in PBS and fixed in 4% paraformaldehyde (PFA) in PBS at 4 °C.
According to a study in Vibrio cholerae[ 51], rpoE mutants attenuate virulence, and the ability of the bacteria to colonise the mouse intestines is reduced.
Indeed, in our perfusion assays, wild-type germ-free mouse intestines were largely tolerant to luminal loading of purified TLR9 agonists, as no transcriptional activation of IL6 and CXCL1 was detected.
Segments of mouse intestine were fixed with 4% formaldehyde in 0.1 M phosphate buffer 20 min at room temperature, followed by overnight incubation at 4°C.
Interestingly, the mouse intestine is capable of producing bioluminescent signal due to processes of digestion.
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