Journal: Experimental & molecular medicine

Article Title: Activation of the osteoblastic HIF-1α pathway partially alleviates the symptoms of STZ-induced type 1 diabetes mellitus via RegIIIγ.

doi: 10.1038/s12276-024-01257-4

Figure Lengend Snippet: Fig. 3 Osteoblastic-derived RegIIIγ assists mice in resisting STZ-induced T1DM. a Representative images of IF staining of RegIIIγ in bone tissue from the RegIIIγ cKO mice and their littermates. b Serum RegIIIγ levels in the RegIIIγ cKO mice and their littermate controls (n = 6). c Random blood glucose levels in the RegIIIγ cKO mice and their littermate controls after STZ induction (n = 5). d Fasting blood glucose levels in the RegIIIγ cKO mice and their littermate controls after STZ induction (n = 6). e Serum insulin levels in the RegIIIγ cKO mice and their littermate controls after STZ induction (n = 5). f GTTs of the RegIIIγ cKO mice and their littermate controls subjected to STZ induction (n = 6). g Compared with the control mice, the RegIIIγ cKO mice exhibited a decreased ability to increase GSIS under STZ conditions. h Representative images of HE-stained pancreatic islets from the RegIIIγ cKO mice and their littermate controls subjected to STZ induction. i, j Representative images of insulin IHC and IF staining of the RegIIIγ cKO mice and their littermate controls subjected to STZ induction. k Insulin mRNA expression in the pancreatic islets of the RegIIIγ cKO mice and their littermate controls after STZ induction (n = 3). l, n Pancreatic islet BAX IHC staining of the RegIIIγ cKO mice and their littermate controls after STZ induction. m, o PCNA IHC staining of pancreatic islets from the RegIIIγ cKO mice and their littermate controls after STZ induction. p The islet equivalent (IEQ) of the RegIIIγ cKO mice was lower than that of their control littermates after STZ treatment (n = 5). q Body weight changes in the RegIIIγ cKO mice and their littermate controls after STZ injection. All data are presented as the mean ± SEM, and p values were analyzed by two-tailed t-tests in (b–e, k, n–p) and two-way ANOVA in (f, g, q). **p < 0.01, ***p < 0.001. All the data are representative of two to three independent experiments. The data were obtained from male mice.

Article Snippet: ELISAs were used to determine the serum insulin (Invitrogen, EMINS), RegIIIγ (Cusabio, CSB-EL019549MO), LCM2 (R&D, MLCN20), and OCN (Abcam, ab285236) levels.

Techniques: Derivative Assay, Staining, Control, Expressing, Immunohistochemistry, Injection, Two Tailed Test

Journal: Experimental & molecular medicine

Article Title: Activation of the osteoblastic HIF-1α pathway partially alleviates the symptoms of STZ-induced type 1 diabetes mellitus via RegIIIγ.

doi: 10.1038/s12276-024-01257-4

Figure Lengend Snippet: Fig. 5 SF-DFO activates the osteoblastic HIF-1α-RegIIIγ pathway and partially alleviates the symptoms of T1DM in adult mice. a WB analysis of HIF-1α and RegIIIγ protein expression in the bone tissues of the mice treated with or without SF-DFO. b HIF-1α and RegIIIγ immunostaining in bone tissue. c, d VEGF and RegIIIγ mRNA expression in the bone tissues of the mice treated with or without SF-DFO (n = 3). e Serum RegIIIγ levels in the mice treated with or without SF-DFO (n = 5). f Fasting blood glucose in the mice treated with STZ/SF-DFO (n = 5). g Serum insulin levels in the mice treated with STZ/SF-DFO (n = 5). h GSIS of the mice treated with STZ/SF-DFO (n = 5). i, k, l Islet HE staining, insulin immunostaining, and insulin, BAX, and PCNA IHC staining images of the mice treated with STZ/SF-DFO (n = 5). j Insulin mRNA expression in the islets of the mice treated with STZ/SF-DFO (n = 3). m Calculation of islet equivalent quantity in the mice treated with STZ/SF- DFO (n = 5). All data are presented as the mean ± SEM, and p values were analyzed by two-tailed t-tests in (c–e); one-way ANOVA in (f, g, j–m); and two-way ANOVA in (h). *p < 0.05, **p < 0.01, ***p < 0.001. All the data are representative of two to three independent experiments. The data were obtained from male mice.

Article Snippet: ELISAs were used to determine the serum insulin (Invitrogen, EMINS), RegIIIγ (Cusabio, CSB-EL019549MO), LCM2 (R&D, MLCN20), and OCN (Abcam, ab285236) levels.

Techniques: Expressing, Immunostaining, Staining, Immunohistochemistry, Two Tailed Test

Journal: Experimental & molecular medicine

Article Title: Activation of the osteoblastic HIF-1α pathway partially alleviates the symptoms of STZ-induced type 1 diabetes mellitus via RegIIIγ.

doi: 10.1038/s12276-024-01257-4

Figure Lengend Snippet: Fig. 6 SF-DFO restores the impaired osteoblastic HIF-1α-RegIIIγ pathway and partially relieves the symptoms of T1DM in elderly mice. a HIF-1α IHC staining of femurs from 8-week-old and 72-week-old young and old mice. b VEGF mRNA expression in bone tissue from the young and old mice after the removal of bone marrow and incubation in an incubator with 1% oxygen for 24 h (n = 6). c Representative images of RegIIIγ IF staining in the young and old mouse femurs. d RegIIIγ mRNA expression in bone tissue from the young and old mice after the removal of bone marrow and incubation in an incubator with 1% oxygen for 24 h (n = 6). e Serum RegIIIγ levels in the old and SF-DFO- treated old mice (n = 6). f Fasting blood glucose in the old and SF-DFO-treated old mice after STZ-induced islet injury (n = 7). g Serum insulin levels in the old and SF-DFO-treated old mice after STZ-induced islet injury (n = 9). h, j, k Representative images of islet HE staining, insulin immunostaining, and BAX and PCNA IHC staining in the old and SF-DFO-treated old mice after STZ-induced islet injury. i Insulin mRNA expression in the islets of the SF-DFO-treated old mice after STZ-induced islet injury. l Calculation of the equivalent islet quantity in the SF- DFO-treated old mice after STZ-induced islet injury. All data are presented as the mean ± SEM, and p values were analyzed by two-tailed t-tests in (e) and one-way ANOVA in (b, d, f, g, i–l). *p < 0.05, **p < 0.01, ***p < 0.001. All the data are representative of two to three independent experiments. The data were obtained from male mice.

Article Snippet: ELISAs were used to determine the serum insulin (Invitrogen, EMINS), RegIIIγ (Cusabio, CSB-EL019549MO), LCM2 (R&D, MLCN20), and OCN (Abcam, ab285236) levels.

Techniques: Immunohistochemistry, Expressing, Incubation, Staining, Immunostaining, Two Tailed Test

Journal: Experimental & molecular medicine

Article Title: Activation of the osteoblastic HIF-1α pathway partially alleviates the symptoms of STZ-induced type 1 diabetes mellitus via RegIIIγ.

doi: 10.1038/s12276-024-01257-4

Figure Lengend Snippet: Fig. 7 Schematic showing how activation of the osteoblastic HIF-1α pathway alleviates the symptoms of STZ-induced T1DM via RegIIIγ. a HIF-1α increases RegIIIγ gene expression by binding to the promoter region of RegIIIγ. b RegIIIγ secreted by osteoblasts enters the blood circulation. c Osteoblast-derived RegIIIγ enters the pancreas through the blood circulation. d Osteoblast-derived RegIIIγ decreases STZ- induced β cell injury. e SF-DFO activates the osteoblastic HIF-1α-RegIIIγ pathway to alleviate STZ-induced pancreatic β cell injury. This scheme was created with BioRender.com.

Article Snippet: ELISAs were used to determine the serum insulin (Invitrogen, EMINS), RegIIIγ (Cusabio, CSB-EL019549MO), LCM2 (R&D, MLCN20), and OCN (Abcam, ab285236) levels.

Techniques: Activation Assay, Gene Expression, Binding Assay, Derivative Assay

Journal: eBioMedicine

Article Title: Intestinal epithelium dysfunctions cause IgA deposition in the kidney glomeruli of intestine-specific Ap1m2 -deficient mice

doi: 10.1016/j.ebiom.2024.105256

Figure Lengend Snippet: Loss of AP-1B decreases IL-22 - STAT3 signal, leading to decreased RegIIIγ expression in the intestine . (A) Western blot analysis of pSTAT3, STAT3, and β-actin in the intestinal epithelium from Ap1m2 fl/fl and Ap1m2 ΔIEC female mice. n = 4. (B) Quantitative PCR analysis of Reg3b and Reg3g in the intestine from ileal Ap1m2 fl/fl and Ap1m2 ΔIEC female mice. Tbp was analyzed as a reference gene. n = 4. (C) Immunofluorescence images of RegIIIγ (red) and nuclei (blue) in the intestine from Ap1m2 fl/fl and Ap1m2 ΔIEC female mice. Scale bars: 100 μm. Data represent the median. ∗ p < 0.05 (unpaired, two-tailed Mann–Whitney test).

Article Snippet: The primary antibodies were as followed: rabbit anti-C3 (1:200, Cat. No. A006302, DAKO, Carpinteria, CA, RRID: AB_578478 ), rat anti-mouse CD107a (LAMP-1) (1:200, clone 1D4B, BioLegend, San Diego, CA, RRID: AB_572020 ), goat anti-mouse IgA (1:200, Cat. No. A90-103A; Betyl Laboratories, RRID: AB_67136 ), rat anti-mouse IgA (1:200, clone RMA-1, BioLegend, RRID: AB_315076 ), Alexa Fluor 488-conjugated donkey anti-mouse IgG (1:400; Cat. No. 715-545-150, Jackson ImmunoResearch, West Grove, PA, RRID: AB_2340846 ), goat anti-mouse pIgR antibody (1:200, Cat. No. AF2800, R&D Systems, Minneapolis, MN, RRID: AB_2283871 ), and rabbit anti-RegIIIγ (1:200, Cat. No. ab198216, Abcam, RRID: AB_2892581 ).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunofluorescence, Two Tailed Test, MANN-WHITNEY