Journal: Scientific Reports

Article Title: Exocrine tissue-driven TFF2 prevents apoptotic cell death of endocrine lineage during pancreas organogenesis

doi: 10.1038/s41598-018-38062-9

Figure Lengend Snippet: Elastase-Cre-mediated Pdx1 inactivation reduces acinar TFF2 in embryonic and neonatal pancreas. ( A ) The expression of TFF2 was detected by RT-PCR in control mice pancreas from E16.5. The original data are shown in Supplementary Fig. . ( B ) Expression of TFF2 is significantly less in Pdx1cKO mice (red) than in control mice (blue). (control mice: n = 7 at E14.5, n = 5 at E16.5, n = 5 at E18.5, and n = 7 at P1; Pdx1cKO mice: n = 5 at E14.5, n = 6 at E16.5, n = 6 at E18.5, and n = 7 at P1; p = N.D at E14.5, p = 0.041 at E16.5, p = 0.0065 at E18.5 and p = 0.0040 at P1). Note that the expression of TFF2 in the mutant stomach is equivalent to that in control stomach at P1 (right panel) (control mice, n = 3, Pdx1cKOmice, n = 3, p = 0.68122). ( C ) Immunostaining of TFF2. TFF2 expression was detected in exocrine cells including the proximal (dotted lines) and distal ducts and acinar cells, but not in islets (arrows) in control mice (upper panels). In Pdx1cKO mice, TFF2 expression was hardly detectable except in the proximal ducts (dotted lines), which were not recombined by Elastase-Cre (bottom panels). These expression patterns were confirmed in at least three individual mice for both genotypes. Scale bars, 100 μm. Bars represent the mean value ± SE. *p < 0.05, **p < 0.01.

Article Snippet: Recombinant mouse TFF2 (#RPA748MU01) was purchased from Uscn Life Science (Texas, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Mutagenesis, Immunostaining

Journal: Scientific Reports

Article Title: Exocrine tissue-driven TFF2 prevents apoptotic cell death of endocrine lineage during pancreas organogenesis

doi: 10.1038/s41598-018-38062-9

Figure Lengend Snippet: TFF2 prevents apoptosis of embryonic insulin-expressing cells through CXCR4. Explant culture of E16.5 pancreatic tissue for 48 hours. ( A ) Immunostaining of insulin (red), beta-catenin (green) and DAPI (blue) for the cultured explant without reagents (left panel), with rTFF2 (middle panel) and with rTFF2 plus AMD3100 (right panel). ( B ) Quantitative analysis of the number of Insulin+ cells. ( C ) Immunostaining of pHH3 (green), insulin (red) and DAPI (blue) for cultured control explant (left panel) and for Pdx1cKO explant without and with rTFF2 (middle and right panels, respectively). The yellow arrowhead shows pHH3-positive Insulin+ cells. ( D ) Quantitative analyses of Insulin+ cell proliferation. ( E ) Immunostaining of TUNEL (green), insulin (red) and DAPI (blue) for cultured Pdx1cKO explant without reagents (left panel), with rTFF2 (middle panel) and with rTFF2 plus AMD3100 (right panel). Yellow arrowheads show TUNEL-positive Insulin+ cells. ( F ) Quantitative analysis of Insulin+ cell apoptosis in Pdx1cKO explant. In control mice pancreas, CXCR4 protected Insulin+ cells from apoptosis. ( G ) Immunostaining of TUNEL (green), insulin (red) and DAPI (blue) for the control explant without and with AMD3100 (left and right panels, respectively). Yellow arrowheads show TUNEL-positive Insulin+ cells. ( H ) Quantitative analyses of Insulin+ cell apoptosis. Scale bars, 100 μm. Bars represent the mean value ± SE. *p < 0.05, **p < 0.01.

Article Snippet: Recombinant mouse TFF2 (#RPA748MU01) was purchased from Uscn Life Science (Texas, USA).

Techniques: Expressing, Immunostaining, Cell Culture, Control, TUNEL Assay

Journal: Scientific Reports

Article Title: Exocrine tissue-driven TFF2 prevents apoptotic cell death of endocrine lineage during pancreas organogenesis

doi: 10.1038/s41598-018-38062-9

Figure Lengend Snippet: Nkx6.1-positive trunk cells are affected by TFF2. Analyses of Nkx6.1+/Insulin- trunk cells in explant culture experiments. ( A ) Immunostaining of Nkx6.1 (green) and insulin (red) for cultured Pdx1cKO explant without reagents (left panel), with rTFF2 (middle panel) and with rTFF2 plus AMD3100 (right panel). ( B ) Quantitative analyses of the number of Nkx6.1+/Insulin- trunk cells. ( C ) TUNEL staining of cultured Pdx1cKO explant without and with rTFF2 (left and right panels, respectively). ( D ) Quantitative analyses of Nkx6.1+/Insulin- trunk cell apoptosis. Scale bars, 100 μm. Bars represent the mean value ± SE. *p < 0.05.

Article Snippet: Recombinant mouse TFF2 (#RPA748MU01) was purchased from Uscn Life Science (Texas, USA).

Techniques: Immunostaining, Cell Culture, TUNEL Assay, Staining