Serca2 deletion in the mouse adult Bmi1+ compartment induces a lethal phenotype involving a severe gastric dysfunction

Figure 3 Impact of conditional Serca2 deletion in the small intestine. A-C: Analysis of crypt-enriched fractions isolated in parallel from DSerca2^Bmi1homo and control animals [5 days post-tamoxifen (Tx)]. A: Estimation of Bmi1⁺ cells (Tmt⁺) by flow cytometry (n = 3 each; independent animals); B: Proliferation analysis (Ki-67⁺) by flow cytometry (n = 3 each; independent animals); C: Evaluation of apoptosis/necrosis rate by flow cytometry (4’,6-diamidino-2-phenylindole/Annexin-V) (n = 3 each; independent animals); D and E: Expression analysis (real-time quantitative polymerase chain reaction) in small intestine segments (duodenum; jejunum; ileum) from DSerca2^Bmi1homo and control mice; Control (n = 5 each; independent animals) and DSerca2^Bmi1homo 5 days (n = 3 each; independent animals) and 7 days (n = 2 each; independent animals) mice. All samples were normalized to levels of the housekeeping gene Hprt. Analysis of a panel of functionally relevant genes in intestinal crypts in crypt-enriched fractions of the indicated animals (n = 3 each; independent animals). D: Analysis of a panel of relevant genes involved in calcium ions regulation in total tissue of the indicated groups of animals (n = 3 each; independent animals); E: Assays were performed three times and data are expressed as mean ± SD. ^aP < 0.05. ^cP < 0.001. FSC: Forward scatter; SSC: Side scatter; DAPI: 4’,6-diamidino-2-phenylindole; Ca²⁺: Calcium ions.