Journal: bioRxiv

Article Title: Genetic background influences the phenotypic penetrance by MAFA S64F MODY in male mice

doi: 10.1101/2025.05.20.653758

Figure Lengend Snippet: A. DEGs identified uniquely in Male Het islets from a mixed background (1238 + 1114 = 2352 genes) versus those uniquely from a C57 background (228 + 855 = 1083 genes) were then overlayed with peaks identified by endogenous MafA CUT&RUN in mouse MIN6 cells (n=11403 peaks). Of these, 250 genes uniquely enriched in a C57 background overlapped with a MafA CUT&RUN peak, while 1210 were uniquely enriched in a Mixed background overlapped with a MafA CUT&RUN peak. B. UCSC Genome Browser tracks showing genomic regions associated with endogenous MafA CUT&RUN peaks near known targets Ins1, Ins2, MafB, and Pdx1, and candidate genes Onecut1, Cry2, Per1, and Per2; MafA CUT&RUN enriched peaks are highlighted in dashed boxes, and regulated genes are depicted below IgG control tracks.

Article Snippet: CUT&RUN was performed on 500,000 dispersed mouse MIN6 cells per condition using CUTANA ChIC/CUT&RUN protocol v3.1 (Epicypher).

Techniques: Control

Journal: bioRxiv

Article Title: Genetic background influences the phenotypic penetrance by MAFA S64F MODY in male mice

doi: 10.1101/2025.05.20.653758

Figure Lengend Snippet: A-B. Immunostaining for MafA show poor detection in Mixed background male Het islets (left) but intact MafA in C57 male Het islets (right). Islets from MafA Λβ included as a negative control. Scale bar, 50μm. C. Left lanes, Western blotting on MIN6 nuclear extract transfected to express either MAFA WT or MAFA S64F shows faster migration in mutant MAFA due to impaired posttranslational modification by phosphorylation. Right lanes, Isolated mouse islets from each genotype and background showed detectable levels of phosphorylated (active) MAFA in C57 background, but relative uniformity of MAFA species with impaired phosphorylation in the Mixed background. D. Quantification of Western blotting bands by Line scan analysis shows greater proportion of phosphorylated MafA species (gray) in Het male islets from the C57 background compared to the Mixed background.

Article Snippet: CUT&RUN was performed on 500,000 dispersed mouse MIN6 cells per condition using CUTANA ChIC/CUT&RUN protocol v3.1 (Epicypher).

Techniques: Immunostaining, Negative Control, Western Blot, Transfection, Migration, Mutagenesis, Modification, Phospho-proteomics, Isolation

Journal: Experimental & Molecular Medicine

Article Title: Mitochondrial-encoded peptide MOTS-c prevents pancreatic islet cell senescence to delay diabetes

doi: 10.1038/s12276-025-01521-1

Figure Lengend Snippet: a , b Treatment with MOTS-c or scrambled control (10 μM, 24 h) and hydrogen peroxide (H 2 O 2 , 200 μM, 24 h) in pancreatic islet cells (pooled from two mice per sample) isolated from littermates of 60-week-old C57BL/6 mice ( n = 3 per sample) led to metabolic changes, as shown in the PCA graph ( a ) and the heat map ( b ). c Enrichment analyses of metabolites were performed for control versus MOTS-c and for H 2 O 2 versus H 2 O 2 + MOTS-c. d A diagram depicting the enriched genes and metabolites analyzed in pancreatic islet cells treated with or without MOTS-c and H 2 O 2 (200 μM, 24 h). e A Venn diagram analysis was performed to find shared pathways by comparing these two enrichment analyses. f Min6 cells overexpressing either empty vector or MOTS-c were treated with glutamine and the expression of genes Slc1a5 , Slc1a5 variant, Gls1/2 and Cd38 , and Cdkn1a and Cdkn2a were assessed. Two-way ANOVA; the error bars are the s.e.m. * P < 0.05, ** P < 0.01 for difference between empty-vector transfected; ## P < 0.01 for difference between empty-vector transfected treated with 5 mM glutamine and MOTS-c transfected treated with 5 mM glutamine. g Min6 cells overexpressing either an empty vector or MOTS-c were analyzed for protein levels of IGF1R, P16, mito-MOTS-c, nuclear MOTS-c- and mTORC1-related molecules and Gls1 in the presence or absence of glutamine (5 mM), H 2 O 2 (200 μM, 24 h) or both.

Article Snippet: All cells except Min6 (ref. ) were purchased from ATCC.

Techniques: Control, Isolation, Plasmid Preparation, Expressing, Variant Assay, Transfection

Journal: Experimental & Molecular Medicine

Article Title: Mitochondrial-encoded peptide MOTS-c prevents pancreatic islet cell senescence to delay diabetes

doi: 10.1038/s12276-025-01521-1

Figure Lengend Snippet: a , b MOTS-c or scrambled ex vivo treatment (10 μM, 24 h) was applied to pancreatic islet cells isolated from four littermates of 60-week-old C57BL/6 mice ( n = 2 per group) to analyze transcriptional changes; these changes were assessed using a PCA plot analyzed by using the scikit-learn Python package ( a ) and a hierarchical heat map ( b ). c – f KEGG and GO analyses (adjusted P value <0.05) indicated that the affected genes are associated with metabolism, cellular communication and signaling and transport ( c ); analysis included: Gene Ontology: biological process (GO: BP) ( d ) Gene Ontology: cellular components (GO: CC) ( e ) Gene Ontology: molecular function (GO: MF) ( f ) were analyzed. g , h The upset plots were used to identify intersecting sets, which are commonly shared genes related to metabolism (pink), signaling (orange) and transport (green); these commonly shared genes, categorized as either upregulated ( g ) or downregulated ( h ) (blue), were displayed in a heat map. i , MOTS-c or scrambled ex vivo treatment (10 μM, 24 h) was applied to pancreatic islet cells isolated from littermates of 60-week-old C57BL/6 mice ( n = 3 per group). j pLJM1-MOTS-c or pLJM1-empty vectors were overexpressed in Min6 cells. Then, cells were treated with or without hydrogen peroxide (200 μM, 24 h) for senescence induction. Subsequently, the expression of genes involved in aspartate–glutamate pathway ( Mdh1 , Mdh1b , Mdh2 , Got1 and Got2 ), EphA5-ephrina5 genes and senescence-related genes ( Cd38 and Grem1 ) were analyzed in both sets.

Article Snippet: All cells except Min6 (ref. ) were purchased from ATCC.

Techniques: Ex Vivo, Isolation, Expressing

Journal: Experimental & Molecular Medicine

Article Title: Mitochondrial-encoded peptide MOTS-c prevents pancreatic islet cell senescence to delay diabetes

doi: 10.1038/s12276-025-01521-1

Figure Lengend Snippet: a Publicly available datasets ( GSE137027 , GSE64553 , GSE72815 , GSE98440 and GSE102004 ) were analyzed for mtRNR1 (MOTS-c) mRNA expression levels (Supplementary Table ). b To explore the underlying mechanism of MOTS-c regulation in senescence, actinonin (50 μM, 24 h) was used to specifically deplete mtDNA in pancreatic islet cells isolated from 12-week-old C57BL/6 mice. c , Min6 cells were transfected with pGenLenti-empty or pGenLenti-Cdkn2a vectors. In b and c the senescence markers (Igf1r, P16INK4a and γ-H2AX) and mTORC1 pathway-related (p-mTOR-2448, p-p70S6K and p-4EBP-1) proteins were analyzed. The pancreatic islet cells from 12- and 90-week-old mice were treated with either hydrogen peroxide (200 μM, 24 h) and doxorubicin (200 nM, 24 h). d The β-gal + p21 + population in pancreatic islets were analyzed. Two-way ANOVA; the error bars are the s.e.m. * P < 0.05, **** P < 0.0001 for comparison. e Hydrogen peroxide and doxorubicin were treated in pancreatic islet cells isolated from 12- or 90-week-old mice to analyze MOTS-c levels. All western blot data are representative of at least three independent experiments. f Treatment with MOTS-c (10 μM, 24 h), with or without H 2 O 2 (200 μM, 24 h), in pancreatic islet cells isolated from 12-week-old C57BL/6 mice prevented senescence markers, including Cdkn1a , Cdkn2a , Cxcl10 and Il-1b mRNA levels. Two-way ANOVA; the error bars are the s.e.m. * P < 0.05, ** P < 0.01, **** P < 0.0001 for comparison. g Pancreatic islet cells isolated from 12-week-old C57BL/6 mice were treated with H 2 O 2 to analyze protein expression levels of γ-H2AX and P16 INK4A . Housekeeping mitochondrial and cytoplasmic proteins (MTCOII and β-actin) were confirmed by western blot. h Treatment with MOTS-c (10 μM, 24 h) in the presence or absence of H 2 O 2 (200 μM, 24 h) in pancreatic islet cells isolated from 12-week-old C57BL/6 mice ( n = 5 per group) was followed by staining and analysis for β-gal, IL-1β, Cxcl10, IL-6 and Igf1r using flow cytometry. Two-way ANOVA; the error bars are the s.e.m. **** P < 0.0001 for comparison.

Article Snippet: All cells except Min6 (ref. ) were purchased from ATCC.

Techniques: Expressing, Isolation, Transfection, Comparison, Western Blot, Staining, Flow Cytometry

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: Taurine supplement alleviates doxorubicin‐induced β‐cell inflammation and senescence. MIN6 cells were pre‐treated with 100 μM taurine for 24 h, followed by 200 nM doxorubicin (DOXO) treatment for 24 h. Cells were cultured in FBS‐free medium to avoid possible contamination of taurine. (A) QPCR analysis of the genes related to inflammation, senescence, and apoptosis in each group of doxorubicin‐induced senescence model. ( n = 3) Relative mRNA levels were normalized to β‐actin. (B) Immunoblotting analysis of p53 and p21 and densitometric quantification. ( n = 3). (C) Immunofluorescence staining of DNA damage marker γ–H2AX in each group (scale bar: 100 μm). ( n = 5). (D) FACS analysis of β‐gal+ PI‐(senescent) and PI+ (dead) MIN6 cells. All results are presented as mean ± SEM. Significance was determined using two‐way ANOVA with Tukey correction. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Cell Culture, Western Blot, Immunofluorescence, Staining, Marker

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: Taurine supplementation alleviates TNF‐α‐induced β‐cell inflammation and senescence. MIN6 cells were pre‐treated with 100 μM taurine for 24 h, followed by 20 ng/mL TNF‐α treatment for 24 h. Cells were cultured in FBS‐free medium to avoid possible contamination of taurine. (A) QPCR analysis of the genes related to senescence in each group of TNF‐α‐induced senescence model. ( n = 4). Relative mRNA levels were normalized to β‐actin. (B) QPCR analysis of the genes related to inflammation and apoptosis in each group. ( n = 4). Relative mRNA levels were normalized to β‐actin. (C) Immunoblotting analysis of p53 and p21 in each group and densitometric quantification. ( n = 3). All results are presented as mean ± SEM. Significance was determined using two‐way ANOVA with Tukey correction. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Cell Culture, Western Blot

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: β‐cells acquire taurine through Slc6a6‐mediated uptake. (A) QPCR analysis of taurine biosynthesis related genes and its transporter Slc6a6 in MIN6 cells and mouse hepatocytes. The results are presented as relative levels over respective gene expression in mouse hepatocytes. ( n = 4). (B, C) MIN6 cells were transfected with siRNA against Scramble or Slc6a6 for 24 h, followed by treatment with taurine (100 μM) or vehicle for 24 h. (B) Immunoblotting analysis of SLC6A6 protein level in each group. ( n = 3). (C) Intracellular taurine levels in the transfected MIN6 cells. ( n = 4). (D) MIN6 cells were pre‐treated with non‐FBS culture medium. The cells were then treated with taurine (100 μM) for 24 h, followed by treatment with SLC6A6 inhibitor (SLC6A6i) (100 μM) or vehicle for 30 min. Intracellular taurine concentration was measured by LC–MS/MS. ( n = 3). All results are presented as mean ± SEM. Significance was determined using two‐tailed independent student's t ‐test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Gene Expression, Transfection, Western Blot, Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Two Tailed Test

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: The protective effects of taurine against β‐cell senescence depend on its transporter SLC6A6. (A, B) MIN6 cells were pre‐treated with the SLC6A6 inhibitor (SLC6A6i) (100 μM) or vehicle for 30 min, followed by treatment with taurine (100 μM) and doxorubicin (200 nM) or vehicle for 24 h in non‐FBS culture medium. The intracellular taurine concentration was then measured by LC–MS/MS. ( n = 3). (B) Immunoblotting analysis of p53 and p21 in each group. (C–F) MIN6 cells were pre‐treated with doxorubicin (200 nM). The cells were then transfected with siRNA against Scramble or Slc6a6 for 24 h, followed by treatment with taurine (100 μM) or vehicle for 24 h. (C) Immunoblotting analysis of SLC6A6, p53, and p21 in each group. ( n = 3). (D) QPCR analysis of gene expressions related to senescence in each group ( n = 4). (E) QPCR analysis of the genes related to β‐cell specific SASP in each group. ( n = 4). (F) QPCR analysis of genes related to inflammation and apoptosis. ( n = 4). All results are presented as mean ± SEM. Significance was determined using two‐way ANOVA with Tukey correction. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Western Blot, Transfection

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: Taurine mitigates senescence, inflammation, and oxidative stress via a p53‐dependent pathway while preserving mitochondrial function independently of p53. (A–C) MIN6 cells were pre‐treated with DOXO (200 nM). The cells were then transfected with siRNA against scramble or p53 for 24 h, followed by treatment with taurine (100 μM) or vehicle for 24 h. Cells were cultured in FBS‐free medium to avoid possible contamination of taurine. (A) QPCR analysis of the genes related to senescence and inflammation in each group. ( n = 4) Relative mRNA levels were normalized to β‐actin. (B) Cellular content of malondialdehyde (MDA) in each group. ( n = 4). (C) Mitochondrial membrane potential was measured using TMRE mitochondrial membrane potential assay. ( n = 7). All results are presented as mean ± SEM. Significance was determined using two‐way ANOVA with Tukey correction. * p < 0.05, ** p < 0.005, *** p < 0.001.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Preserving, Transfection, Cell Culture, Membrane

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: Identification of Taurine‐CDKN2AIP binding in pancreatic β cells. (A, B) Limited proteolysis‐mass spectrometry (LiP‐MS) was used to screen for taurine interacting proteins in the INS1E β‐cell proteome. Heatmap shows potential taurine binding targets identified by LiP‐MS. Vehicle: N = 3. Taurine: N = 3. (B) p53 pathway related proteins levels between two groups and their binding scores with taurine. (C) Three‐dimensional diagram of the binding modes between human CDKN2AIP and taurine. Taurine potentially binds to CDKN2AIP via residues PRO484, LEU485, LYS486. (D) DARTS analysis using MIN6 cell lysates incubated with taurine. (E) DARTS analysis using INS1E cell lysates incubated with taurine. (F) 500 ng of CDKN2AIP recombinant protein were subjected to SDS‐PAGE and silver staining to assess purity. (G) DARTS analysis using CDKN2AIP recombinant protein incubated with taurine. (H) MIN6 cells treated with taurine (100 μM, 24 h) or vehicle were subjected to immunoprecipitation against CDKN2AIP.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Binding Assay, Mass Spectrometry, Incubation, Recombinant, SDS Page, Silver Staining, Immunoprecipitation

Journal: Journal of Diabetes

Article Title: Taurine Alleviates Pancreatic β‐Cell Senescence by Inhibition of p53 Pathway

doi: 10.1111/1753-0407.70100

Figure Lengend Snippet: Taurine treatment accelerates p53 degradation by binding to CDKN2AIP. (A) p53 protein degradation was detected using cycloheximide (CHX, 10 μM) chase assay. (B) HEK 293 cells were transfected with plasmids encoding GFP‐tagged CDKN2AIP (WT) and CDKN2AIP‐triple mutant (MT) for 48 h. DARTS analysis was performed using cell lysates incubated with taurine, followed by immunoblotting analysis as indicated. (C) INS‐1E cells were transfected with plasmids encoding GFP control, GFP‐tagged CDKN2AIP, and its triple mutant for 24 h, followed by taurine treatment for 24 h. Immunoblotting analysis of CDKN2AIP and p53 in each group.

Article Snippet: Mouse pancreatic β‐cell line MIN6 (AddexBio Technologies, Cat#C0018008) and rat insulinoma cell INS‐1E (AddexBio Technologies, Cat#C0018009) were cultured in DMEM (Gibco, Cat#12800082) or RPMI 1640 supplemented with 15% FBS, 1% penicillin–streptomycin, and 50 μM β‐mercaptoethanol.

Techniques: Binding Assay, Transfection, Mutagenesis, Incubation, Western Blot, Control

Journal: Cell Reports Medicine

Article Title: RSPO1, a potent inducer of pancreatic β cell neogenesis

doi: 10.1016/j.xcrm.2025.102126

Figure Lengend Snippet: RSPO1 induces β cell replication in MIN6 cells, in isolated islets, and in adult WT mice (A) Assessment of MIN6 cell number upon 24-h incubation with increasing doses of native RSPO1 or saline. (B) Representative photographs of pancreatic islets isolated from WT adult mice incubated with either RSPO1 (at 0.4, 1, or 2 μM) or saline. Aiming to label replicating cells, isolated islets were co-incubated with BrdU during the last 24 h and then stained for insulin (red) and BrdU (green). (C) Quantification of the percentage of BrdU + β cells in control islets and islets incubated for 72 h with increasing doses of RSPO1. (D) Pancreatic sections obtained from adult WT mice administered intraperitoneally for 5 consecutive days with different doses of RSPO1 and stained for insulin (red) and Ki67 (green). (E) Quantitative assessment of Ki67 + cells per islet after 5 consecutive RSPO1 administrations. All data shown represent mean ± SEM of n = 5. Results were considered significant if p < 0.0001 (∗∗∗∗), p < 0.001 (∗∗∗), p < 0.01 (∗∗), and p < 0.05 (∗) using one-way ANOVA. See also .

Article Snippet: The pancreatic immortalized β-cell lines MIN6 (AddexBio) and Ins1 (ATCC), the α-cell-derived αTC1 (ATCC) and the acinar cell line 266-6 (ATCC) were cultured in 100mm Petri dishes (Eppendorf) in cell line-specific culture media indicated by manufacturer’s instructions and maintained in an humified incubator at 37°C and 5% CO2.

Techniques: Isolation, Incubation, Saline, Staining, Control, IF-P

Journal: Cell Reports Medicine

Article Title: RSPO1, a potent inducer of pancreatic β cell neogenesis

doi: 10.1016/j.xcrm.2025.102126

Figure Lengend Snippet: RSPO1 stimulates β cell replication via the activation of Wnt signaling The putative activation of the canonical Wnt signaling pathway upon RSPO1 incubation was assessed in vitro . (A) MIN6 cell number quantification upon incubation for 24 h with increasing doses of two RSPO1 mutants Lgr4 M and Znrf3 M . (B) MIN6 cell number assessment upon a 24-h treatment with saline, 0.1% DMSO, 100 nM MSAB, or 500 nM cardamonin and co-incubation of MIN6 cells with either MSAB or cardamonin and native RSPO1 at 400 nM. (C) Quantification of BrdU + cells in ex vivo murine islets incubated for 72 h with saline, 100 nM MSAB, 1 μM RSPO1, or a combination of the 2. (D) β-catenin protein levels upon RSPO1 treatment assessed by ELISA following a 3-h incubation with saline, native RSPO1, RSPO1 Lgr4 M , or RSPO1 Znrf3 M at 400 nM. Data shown represent mean ± SEM of n = 5. Results were considered significant if p < 0.0001 (∗∗∗∗), p < 0.001 (∗∗∗), p < 0.01 (∗∗), and p < 0.05 (∗) using one-way ANOVA. See also .

Article Snippet: The pancreatic immortalized β-cell lines MIN6 (AddexBio) and Ins1 (ATCC), the α-cell-derived αTC1 (ATCC) and the acinar cell line 266-6 (ATCC) were cultured in 100mm Petri dishes (Eppendorf) in cell line-specific culture media indicated by manufacturer’s instructions and maintained in an humified incubator at 37°C and 5% CO2.

Techniques: Activation Assay, Incubation, In Vitro, Saline, Ex Vivo, Enzyme-linked Immunosorbent Assay, IF-P

Journal: Cell Reports Medicine

Article Title: RSPO1, a potent inducer of pancreatic β cell neogenesis

doi: 10.1016/j.xcrm.2025.102126

Figure Lengend Snippet: An FC-coupled RSPO1 induces pancreatic β cell neogenesis in vitro , ex vivo , and in vivo (A) Assessment of MIN6 cell number upon 24-h incubation with saline or increasing doses of FC-coupled RSPO1 protein ( n = 5). (B) Quantification of the percentage of BrdU+ cells in saline-treated islets and islets incubated for 72 h with 200 nM or 1 or 3 μM FC-RSPO1 ( n = 5). (C) Weekly monitoring of random glycemia in 10-week-old NOD females injected intraperitoneally weekly for 18 weeks with either saline or 2.4 mg/kg of FC-RSPO1 ( n = 12 in control group, n = 10 in treated group). (D) Quantification of the whole β cell mass in NOD mice weekly administered with either saline or 2.4 mg/kg of FC-coupled RSPO1. The β cell mass of 10-week-old NOD females was used to evaluate the insulin + area at the beginning of the study ( n = 12 in control group, n = 10 in treated group). (E–H) Pancreatic sections from FC-RSPO1-treated NOD mice stained for the β cell markers Pdx1 (E), PC1/3 (F), Nkx6.1 (G), and Glut2 (H). All data shown represent mean ± SEM. Results were considered significant if p < 0.0001 (∗∗∗∗), p < 0.001 (∗∗∗), p < 0.01 (∗∗), and p < 0.05 (∗) following a one-way ANOVA (A and B); a one-way ANOVA, a Mann-Whitney test, or a Kruskal-Wallis test (C); or an unpaired Student’s t test (D).

Article Snippet: The pancreatic immortalized β-cell lines MIN6 (AddexBio) and Ins1 (ATCC), the α-cell-derived αTC1 (ATCC) and the acinar cell line 266-6 (ATCC) were cultured in 100mm Petri dishes (Eppendorf) in cell line-specific culture media indicated by manufacturer’s instructions and maintained in an humified incubator at 37°C and 5% CO2.

Techniques: In Vitro, Ex Vivo, In Vivo, Incubation, Saline, Injection, Control, Staining, IF-P, MANN-WHITNEY