Journal: Advanced Science

Article Title: Beta‐Cell Tipe1 Orchestrates Insulin Secretion and Cell Proliferation by Promoting Gαs/cAMP Signaling via USP5

doi: 10.1002/advs.202304940

Figure Lengend Snippet: Sequences of real‐time PCR primers.

Article Snippet: Pdx1‐Cre mice (Stock No. 01 4647) were purchased from Cyagen Biosciences Inc. Tipe1‐Loxp ( Tipe1 f/f ) mice were generated by Biocytogen Corporation (Beijing, China), as described previously.

Techniques: Real-time Polymerase Chain Reaction

Journal: Cell metabolism

Article Title: Impaired Islet Function in Commonly Used Transgenic Mouse Lines due to Human Growth Hormone Minigene Expression

doi: 10.1016/j.cmet.2014.11.004

Figure Lengend Snippet: β Cell-Specific Transgenic Mice Generated Based on a Similar Strategy, Namely Using the hGH Minigene as Transgene Enhancer

Article Snippet: Pdx1-Cre Late transgenic mice ( Herrera, 2000 ) (Dr. Herrera, University of Geneva, Switzerland) were crossed with C57BL/6J (Janvier) for at least eight generations.

Techniques: Transgenic Assay, Generated

Journal: Diabetologia

Article Title: XBP1 maintains beta cell identity, represses beta-to-alpha cell transdifferentiation and protects against diabetic beta cell failure during metabolic stress in mice

doi: 10.1007/s00125-022-05669-7

Figure Lengend Snippet: Beta cell-specific Xbp1 deletion in adult mice fed a high-fat diet triggers diabetes by disrupting insulin secretory capacity. ( a ) Fed blood glucose levels. ( b ) Blood glucose levels and AUC for glucose during OGTT. ( c ) Serum insulin levels and AUC for insulin during OGTT. ( d ) Insulin secretion from isolated islets treated with low (2 mmol/l) or high (20 mmol/l) stimulatory level of glucose. ( e ) Insulin content in islets. ( f ) mRNA expression of insulin and Pc2 in islets expressed as fold change of the levels in chow-fed β-Xbp1 +/+ mice. ( g ) Serum proinsulin levels. ( h ) mRNA expression of adaptive UPR genes in islets expressed as fold change of the levels in chow-fed β-Xbp1 +/+ mice. ( i ) mRNA expression of pro-apoptosis UPR genes in islets expressed as fold change of the levels in chow-fed β-Xbp1 +/+ mice. ( j ) mRNA expression of beta cell function genes in islets expressed as fold change of the levels in chow-fed β-Xbp1 +/+ mice. All data are represented as means ± SEM. n = 5–11, chow-fed β-Xbp1 +/+ ; n = 6–19, high-fat-fed β-Xbp1 +/+ ; n = 5–16, chow-fed β-Xbp1 −/− ; n = 6–18, high-fat-fed β-Xbp1 −/− . ANOVA: * p <0.05, ** p <0.01, *** p <0.001 genotype effect; † p <0.05, †† p <0.01, ††† p <0.001 diet effect. C, chow-fed; HF, high-fat-fed; Tam., tamoxifen

Article Snippet: A model for cell type-specific lineage tracing and transcript profiling was developed by crossing Xbp1 flox/flox Pdx1 -Cre ER mice with Gt/Rosa26 GFP [ Gt(ROSA)26Sor t m9(EGFP/Rpl10a)Amc ; The Jackson Laboratory] mice.

Techniques: Isolation, Expressing, Cell Function Assay

Journal: Diabetologia

Article Title: XBP1 maintains beta cell identity, represses beta-to-alpha cell transdifferentiation and protects against diabetic beta cell failure during metabolic stress in mice

doi: 10.1007/s00125-022-05669-7

Figure Lengend Snippet: Xbp1 deletion leads to altered islet cell composition, increased beta cell turnover, beta cell dedifferentiation and beta-to-alpha cell transdifferentiation. ( a ) Pancreas weight. ( b ) Beta cell mass (quantification of immunostaining for insulin). ( c ) Beta cell proliferation rate (quantification of immunostaining for Ki-67 and insulin). ( d ) Beta cell apoptosis rate (quantification of immunostaining for TUNEL and insulin). ( e ) Representative images of immunostaining for insulin and glucagon. ( f ) Alpha cell mass (quantification of immunostaining for glucagon). ( g ) mRNA expression of genes involved in beta cell proliferation ( Myc ), dedifferentiation ( Sox9 ) and senescence ( p21 and p53 ) in islets, expressed as fold change of the levels in chow-fed β-Xbp1 +/+ mice. ( h ) Representative images of immunostaining for GFP (green), glucagon (red) and insulin (magenta). Scale bar, 20 μm. ( i ) Expression in immunoprecipitated mRNA of beta cell identity ( Pdx1 , Beta2 , Nkx6.1 and Foxo1 ) genes expressed as fold change of the levels in β-Xbp1 +/+ Gt mice, and beta cell dedifferentiation ( Aldh1a3 ) and alpha cell ( Arx , Irx2 and Gcg ) genes expressed as fold change of the levels in β-Xbp1 −/− Gt mice. All data are represented as means ± SEM. ( a – g ) n = 4–8, chow-fed β-Xbp1 +/+ ; n = 6–10, high-fat-fed β-Xbp1 +/+ ; n = 4–7, chow-fed β-Xbp1 −/− ; n = 6–9, high-fat-fed β-Xbp1 −/− . ANOVA: * p <0.05, ** p <0.01, *** p <0.001 genotype effect; † p <0.05, †† p <0.01 diet effect. ( h , i ) n = 3–7, β-Xbp1 +/+ Gt; n = 3–8, β-Xbp1 −/− Gt. Unpaired two-tailed t test: * p <0.05, ** p <0.01, *** p <0.001. C, chow-fed; HF, high-fat-fed. Gluc, glucagon; Ins, insulin

Article Snippet: A model for cell type-specific lineage tracing and transcript profiling was developed by crossing Xbp1 flox/flox Pdx1 -Cre ER mice with Gt/Rosa26 GFP [ Gt(ROSA)26Sor t m9(EGFP/Rpl10a)Amc ; The Jackson Laboratory] mice.

Techniques: Immunostaining, TUNEL Assay, Expressing, Immunoprecipitation, Two Tailed Test

Journal: Diabetologia

Article Title: XBP1 maintains beta cell identity, represses beta-to-alpha cell transdifferentiation and protects against diabetic beta cell failure during metabolic stress in mice

doi: 10.1007/s00125-022-05669-7

Figure Lengend Snippet: Beta cell-specific Xbp1 deletion in obese o b/ob mice leads to diabetes due to failure of beta cell compensation. ( a ) Fed blood glucose levels. ( b ) Blood glucose levels and AUC for glucose during OGTT. ( c ) Serum insulin levels and AUC for insulin during OGTT. ( d ) mRNA expression of adaptive UPR genes in islets expressed as fold change of the levels in β-Xbp1 +/+ Wt mice. ( e ) mRNA expression of pro-apoptosis UPR genes in islets expressed as fold change of the levels in β-Xbp1 +/+ Wt mice. ( f ) mRNA expression of insulin and Pc2 in islets expressed as fold change of the levels in β-Xbp1 +/+ Wt mice. ( g ) Serum proinsulin levels. ( h ) Insulin secretion from isolated islets treated with low (2 mmol/l) or high (20 mmol/l) stimulatory level of glucose for 1 h. ( i ) Insulin content in islets. ( j ) Insulin secretion from isolated islets of β-Xbp1 +/+ Ob and β-Xbp1 −/− Ob mice treated with repeated low (2 mmol/l) or high (20 mmol/l) stimulatory level of glucose for three cycles. ( k ) Insulin content in islets following three cycles of glucose treatment. ( l ) Representative images of TUNEL immunostaining in pancreas sections. ( m ) Beta cell apoptosis rate (quantification of immunostaining for TUNEL and insulin). All data are represented as means ± SEM. ( a – i ) n = 3–12, β-Xbp1 +/+ Wt; n = 3–14, β-Xbp1 −/− Wt; n = 3–7, β-Xbp1 +/+ Ob; n = 7–13, β-Xbp1 −/− Ob. ( j , k ) n = 5, β-Xbp1 +/+ Ob; n = 4, β-Xbp1 −/− Ob. ( l , m ) n = 6, β-Xbp1 +/+ Wt; n = 6, β-Xbp1 −/− Wt; n = 6, β-Xbp1 +/+ Ob; n = 8, β-Xbp1 −/− Ob. ANOVA: * p <0.05, ** p <0.01, *** p <0.001 Xbp1 genotype effect; † p <0.05, †† p <0.01, ††† p <0.001 Ob genotype effect. Tam., tamoxifen

Article Snippet: A model for cell type-specific lineage tracing and transcript profiling was developed by crossing Xbp1 flox/flox Pdx1 -Cre ER mice with Gt/Rosa26 GFP [ Gt(ROSA)26Sor t m9(EGFP/Rpl10a)Amc ; The Jackson Laboratory] mice.

Techniques: Expressing, Isolation, TUNEL Assay, Immunostaining

Journal: Diabetologia

Article Title: XBP1 maintains beta cell identity, represses beta-to-alpha cell transdifferentiation and protects against diabetic beta cell failure during metabolic stress in mice

doi: 10.1007/s00125-022-05669-7

Figure Lengend Snippet: Xbp1 deletion in beta cells potentiates glucolipoapoptosis by attenuating the antioxidant response. ( a ) Apoptosis rate in islets determined by cell death detection ELISA. ( b ) mRNA expression of adaptive UPR genes in islets expressed as fold change of the levels in β-Xbp1 +/+ islets. ( c ) mRNA expression of pro-apoptosis UPR genes in islets expressed as fold change of the levels in β-Xbp1 +/+ islets. ( d ) mRNA expression of antioxidant enzymes in islets expressed as fold change of the levels in β-Xbp1 +/+ islets. ( e ) Apoptosis rate in islets co-treated in the absence or presence of the antioxidant. Islets were treated in the absence (control, C) or presence of high GP for 72 h. All data are represented as means±SEM. n = 6–9, β-Xbp1 +/+ ; n = 8–9, β-Xbp1 −/− . ANOVA: * p <0.05, ** p <0.01, *** p <0.001 genotype effect; † p <0.05, †† p <0.01, ††† p <0.001 treatment effect

Article Snippet: A model for cell type-specific lineage tracing and transcript profiling was developed by crossing Xbp1 flox/flox Pdx1 -Cre ER mice with Gt/Rosa26 GFP [ Gt(ROSA)26Sor t m9(EGFP/Rpl10a)Amc ; The Jackson Laboratory] mice.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Control