Journal: Frontiers in Nutrition

Article Title: High-fat diet mouse model receiving L-glucose supplementations propagates liver injury

doi: 10.3389/fnut.2024.1469952

Figure Lengend Snippet: Metabolic profile assessment: (A) Fasting blood sugar (FBS) was assessed as indicated in materials and methods. GLUT2 expressions were quantitated by (B) western blot and (C) RT-PCR methods. (D) Serum insulin C-peptide measured by ELISA. (E) Hepatic expression of inulin receptor (IR) was evaluated from liver sections by western blot (F) HOMA2 calculator calculated HOMA-IR. Paired and unpaired Student’s t -test and ANOVA were used for statistically significant differences. p value was compared between RD and HFD control groups or within RD or HFD groups. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Rabbit anti-Human/Mouse GYS2 (Proteintech, 22371-1-AP), Rabbit anti-Human/Mouse Glycogen synthase [p Ser641] (Novus bio, NBP2-67315), rabbit anti-human/mouse PYGL antibody (Proteintech, 15851-1-AP), rabbit anti-human/mouse Glut2 polyclonal antibody (Proteintech, 20436-1-AP), rabbit anti-human/mouse ADRP/Perilipin 2 Polyclonal antibody (Proteintech, 15294-1-AP), rabbit anti-human/mouse Alpha Smooth Muscle antibody (Novus, NBP1-30894), mice anti-human/mouse AKT antibody (R&D, MAB 2055), mice anti-human/mouse phospho-AKT antibody (R&D, MAB 887), rabbit anti-human/mouse Insulin Receptor-beta antibody (Proteintech, 20433-1-AP), and rabbit anti-human/mouse beta Actin polyclonal antibody (Proteintech, 20536-1-AP).

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Increase in the Expression of Glucose Transporter 2 (GLUT2) on the Peripheral Blood Insulin-Producing Cells (PB-IPC) in Type 1 Diabetic Patients after Receiving Stem Cell Educator Therapy

doi: 10.3390/ijms25158337

Figure Lengend Snippet: Phenotypic characterization of PB-IPC in human peripheral blood. ( A ) Expression of GLUT2 + CD45RO + PB-IPC in healthy donors. Representative data from one of twelve preparations. Flow cytometry analysis showed the gating strategy by using the FITC-conjugated anti-human lineage cocktail 1 (Lin1) (CD3, CD14, CD16, CD19, CD20, CD56) to eliminate the known cellular lineages such as T cells, monocytes/macrophages, granulocytes, B cells, and natural killer (NK) cells. Anti-human leukocyte common antigen CD45 was utilized to exclude the red blood cells (RBC) and platelets’ contamination during data analysis. CD34 was applied to discriminate the hematopoietic stem cells. Isotype-matched IgGs served as negative controls. ( B ) Low percentage of GLUT2 + CD45RO + PB-IPC in recent onset T1D patients (n = 9) in comparison with the healthy control (n = 12).

Article Snippet: Phenotypic characterization of PB-IPC and IL-1β-positive cells was performed by flow cytometry with specific markers including FITC-conjugated anti-human lineage cocktail 1 (Lin1) (CD3, CD14, CD16, CD19, CD20, CD56), PE-conjugated anti-human GLUT2 antibody (R & D Systems, Minneapolis, MN, USA), phycoerythrin-Cy5.5 (PE-Cy5.5)-conjugated mouse anti-human SOX2 (Biolegend, San Diego, CA, USA), PE-Cy7-conjugated mouse anti-human CD45RO (Biolegend, San Diego, CA, USA), APC-Alexa Fluor 750-conjugated mouse anti-human CCR7 (Biolegend, San Diego, CA, USA), Alexa Fluor 647-conjugated mouse anti-human OCT3/4 (Biolegend, San Diego, CA, USA), BV421-conjugated mouse anti-human CD34 (Biolegend, San Diego, CA, USA), BV510-conjugated mouse anti-human leukocyte common antigen CD45 (Biolegend, San Diego, CA, USA), pacific blue (PB)-conjugated mouse anti-human IL-1β (Biolegend, San Diego, CA, USA), FITC-conjugated mouse anti-human interferon γ (Beckman Coulter, Brea, CA, USA), FITC-conjugated mouse anti-human interferon γ (Beckman Coulter, Brea, CA, USA), PE Cy7-conjugated mouse anti-human IL-13 (Biolegend, San Diego, CA, USA), APC-conjugated mouse anti-human IL-17A (Biolegend, San Diego, CA, USA), and pacific blue (PB)-conjugated mouse anti-human tumor necrosis factor (TNF)α (Biolegend, San Diego, CA, USA).

Techniques: Expressing, Flow Cytometry, Comparison, Control

Journal: International Journal of Molecular Sciences

Article Title: Increase in the Expression of Glucose Transporter 2 (GLUT2) on the Peripheral Blood Insulin-Producing Cells (PB-IPC) in Type 1 Diabetic Patients after Receiving Stem Cell Educator Therapy

doi: 10.3390/ijms25158337

Figure Lengend Snippet: Increase in the percentage of GLUT2 + CD45RO + PB-IPC in patients after the treatment with stem cell educator therapy. Patients received the stem cell educator therapy according to the FDA-approved clinical protocol (IND #19247) for type 1 diabetes. Flow cytometry analysis was performed at the baseline before and after receiving stem cell educator therapy. The GLUT2 + CD45RO + PB-IPC were gated from the population of Lin1 − CD34 − CD45 + CD45RO + CCR7 + cells. Isotype-matched IgGs served as negative controls. ( A ) The percentage of GLUT2 + CD45RO + PB-IPC was upregulated in T1D subjects one month after stem cell educator therapy. n = 10. The Wilcoxon matched-pairs signed rank test was utilized for the non-parametric data. ( B ) Representative data showed the increased expressions of GLUT2 + CD45RO + PB-IPC in T1D patient HU2058 before and after the stem cell educator therapy. ( C ) Representative data showed the increased expressions of GLUT2 + CD45RO + PB-IPC in a prediabetic patient HU2039 before and after the stem cell educator therapy.

Article Snippet: Phenotypic characterization of PB-IPC and IL-1β-positive cells was performed by flow cytometry with specific markers including FITC-conjugated anti-human lineage cocktail 1 (Lin1) (CD3, CD14, CD16, CD19, CD20, CD56), PE-conjugated anti-human GLUT2 antibody (R & D Systems, Minneapolis, MN, USA), phycoerythrin-Cy5.5 (PE-Cy5.5)-conjugated mouse anti-human SOX2 (Biolegend, San Diego, CA, USA), PE-Cy7-conjugated mouse anti-human CD45RO (Biolegend, San Diego, CA, USA), APC-Alexa Fluor 750-conjugated mouse anti-human CCR7 (Biolegend, San Diego, CA, USA), Alexa Fluor 647-conjugated mouse anti-human OCT3/4 (Biolegend, San Diego, CA, USA), BV421-conjugated mouse anti-human CD34 (Biolegend, San Diego, CA, USA), BV510-conjugated mouse anti-human leukocyte common antigen CD45 (Biolegend, San Diego, CA, USA), pacific blue (PB)-conjugated mouse anti-human IL-1β (Biolegend, San Diego, CA, USA), FITC-conjugated mouse anti-human interferon γ (Beckman Coulter, Brea, CA, USA), FITC-conjugated mouse anti-human interferon γ (Beckman Coulter, Brea, CA, USA), PE Cy7-conjugated mouse anti-human IL-13 (Biolegend, San Diego, CA, USA), APC-conjugated mouse anti-human IL-17A (Biolegend, San Diego, CA, USA), and pacific blue (PB)-conjugated mouse anti-human tumor necrosis factor (TNF)α (Biolegend, San Diego, CA, USA).

Techniques: Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Increase in the Expression of Glucose Transporter 2 (GLUT2) on the Peripheral Blood Insulin-Producing Cells (PB-IPC) in Type 1 Diabetic Patients after Receiving Stem Cell Educator Therapy

doi: 10.3390/ijms25158337

Figure Lengend Snippet: Increase in the percentage of GLUT2 + CD45RO + PB-IPC in patients with longstanding T1D after the treatment with stem cell educator therapy. ( A ) Markedly upregulated percentages of GLUT2 + CD45RO + PB-IPC in patients (n = 8) with longstanding T1D after the treatment with stem cell educator therapy for 1 month. ( B ) Representative data from subject HU2003 (diagnosed with T1D for 58 years since age 1 year old) showing improved expression of GLUT2 + CD45RO + PB-IPC and continued increased level of GLUT2 + CD45RO + PB-IPC at the twelfth month follow-up.

Article Snippet: Phenotypic characterization of PB-IPC and IL-1β-positive cells was performed by flow cytometry with specific markers including FITC-conjugated anti-human lineage cocktail 1 (Lin1) (CD3, CD14, CD16, CD19, CD20, CD56), PE-conjugated anti-human GLUT2 antibody (R & D Systems, Minneapolis, MN, USA), phycoerythrin-Cy5.5 (PE-Cy5.5)-conjugated mouse anti-human SOX2 (Biolegend, San Diego, CA, USA), PE-Cy7-conjugated mouse anti-human CD45RO (Biolegend, San Diego, CA, USA), APC-Alexa Fluor 750-conjugated mouse anti-human CCR7 (Biolegend, San Diego, CA, USA), Alexa Fluor 647-conjugated mouse anti-human OCT3/4 (Biolegend, San Diego, CA, USA), BV421-conjugated mouse anti-human CD34 (Biolegend, San Diego, CA, USA), BV510-conjugated mouse anti-human leukocyte common antigen CD45 (Biolegend, San Diego, CA, USA), pacific blue (PB)-conjugated mouse anti-human IL-1β (Biolegend, San Diego, CA, USA), FITC-conjugated mouse anti-human interferon γ (Beckman Coulter, Brea, CA, USA), FITC-conjugated mouse anti-human interferon γ (Beckman Coulter, Brea, CA, USA), PE Cy7-conjugated mouse anti-human IL-13 (Biolegend, San Diego, CA, USA), APC-conjugated mouse anti-human IL-17A (Biolegend, San Diego, CA, USA), and pacific blue (PB)-conjugated mouse anti-human tumor necrosis factor (TNF)α (Biolegend, San Diego, CA, USA).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Increase in the Expression of Glucose Transporter 2 (GLUT2) on the Peripheral Blood Insulin-Producing Cells (PB-IPC) in Type 1 Diabetic Patients after Receiving Stem Cell Educator Therapy

doi: 10.3390/ijms25158337

Figure Lengend Snippet: Increased expression of GLUT2 on PB-IPC in other patients after receiving stem cell educator therapy. ( A ) Improvement of the GLUT2 expression on PB-IPC of patients with alopecia areata (AA, n = 8.) after stem cell educator therapy for 3 months. ( B ) Upregulation of the GLUT2 expression on PB-IPC of patients with autoimmune- or inflammation-associated diseases (n = 8) after stem cell educator therapy for 1 month. ( C ) Flow cytometry analysis showing the increased percentages of GLUT2 expression from the baseline levels (left panels) to normal levels post-treatment (right panels) with stem cell educator therapy. Representative data were obtained from subjects HU2027 with AA (top panels), HU2054 with Parkinson’s disease (middle panels), and HU2064 with longstanding Eczema (bottom panels), respectively.

Article Snippet: Phenotypic characterization of PB-IPC and IL-1β-positive cells was performed by flow cytometry with specific markers including FITC-conjugated anti-human lineage cocktail 1 (Lin1) (CD3, CD14, CD16, CD19, CD20, CD56), PE-conjugated anti-human GLUT2 antibody (R & D Systems, Minneapolis, MN, USA), phycoerythrin-Cy5.5 (PE-Cy5.5)-conjugated mouse anti-human SOX2 (Biolegend, San Diego, CA, USA), PE-Cy7-conjugated mouse anti-human CD45RO (Biolegend, San Diego, CA, USA), APC-Alexa Fluor 750-conjugated mouse anti-human CCR7 (Biolegend, San Diego, CA, USA), Alexa Fluor 647-conjugated mouse anti-human OCT3/4 (Biolegend, San Diego, CA, USA), BV421-conjugated mouse anti-human CD34 (Biolegend, San Diego, CA, USA), BV510-conjugated mouse anti-human leukocyte common antigen CD45 (Biolegend, San Diego, CA, USA), pacific blue (PB)-conjugated mouse anti-human IL-1β (Biolegend, San Diego, CA, USA), FITC-conjugated mouse anti-human interferon γ (Beckman Coulter, Brea, CA, USA), FITC-conjugated mouse anti-human interferon γ (Beckman Coulter, Brea, CA, USA), PE Cy7-conjugated mouse anti-human IL-13 (Biolegend, San Diego, CA, USA), APC-conjugated mouse anti-human IL-17A (Biolegend, San Diego, CA, USA), and pacific blue (PB)-conjugated mouse anti-human tumor necrosis factor (TNF)α (Biolegend, San Diego, CA, USA).

Techniques: Expressing, Flow Cytometry

Journal: International Journal of Biological Sciences

Article Title: Intracellular Insulin and Impaired Autophagy in a Zebrafish model and a Cell Model of Type 2 diabetes

doi: 10.7150/ijbs.19249

Figure Lengend Snippet: Long-term high-fat diet induced Type 2 Diabetes-like symptoms in zebrafish. Adult zebrafish were fed a normal diet (Ctrl) or a high-fat diet (HFD) for 10 weeks. (A)-(C) Comparison of morphology (A), body weight (B), and body mass index (BMI; weight/length, g/cm) (C) between Ctrl and HFD zebrafish. (D) Higher fasting blood glucose induced in the HFD group compared to the normal diet group. (E) Representative images of hematoxylin and eosin (H&E) staining show lipid vesicles in zebrafish liver slices in both groups. The histogram shows the average number of fat cavities per sight field in the control group and HFD group, which were counted from ten sight fields each group. (F) Representative electrophorogram of insulin bands from RT-PCR (left) and real-time qPCR results (right) show the mRNA levels of insulin gene transcription in the liver, brain, and muscle of zebrafish in both groups. (G) The mRNA levels of insulin receptor substrate-2 (IRS-2) and glucose transporter 2 (GLUT2) assayed by RT-PCR electrophorogram (left) and real-time qPCR results (right) in zebrafish liver. *p<0.05, **p<0.01, ***p<0.001 vs the control group.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibody including anti-human p62 polyclonal antibody (PM045, MBL, Nagoya, Japan) diluted 1:500, and anti-human β-ACTIN monoclonal antibody (TA-09, ZSGB-Bio, Beijing, China) diluted 1:1000, anti-human LC3B polyclonal antibody (PM036, MBL) diluted 1:500, anti-human LAMP2 monoclonal antibody (sc-5571, Santa Cruz Biotechnology, Dallas, TX) diluted 1:200, anti-human PPARA monoclonal antibody (AM8452b, Abgent, Suzhou, China) diluted 1:500, anti-human PGC-1α polyclonal antibody (ab54481, Abcam, Cambridge, UK) diluted 1:500, anti-human insulin polyclonal antibody (sc-7838,Santa Cruz Biotechnology) diluted 1:200, anti-human GLUT2 polyclonal antibody (sc-9117,Santa Cruz Biotechnology) diluted 1:200, and anti-human IRS2 polyclonal antibody (#4502, CST, Danvers, MA, USA) diluted 1:200.

Techniques: Staining, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Biological Sciences

Article Title: Intracellular Insulin and Impaired Autophagy in a Zebrafish model and a Cell Model of Type 2 diabetes

doi: 10.7150/ijbs.19249

Figure Lengend Snippet: Palmitic acid induced type 2 diabetes-like features in HepG2 cells. (A) Lipid level was increased in PA concentration-dependent manner in PA-treated cells determined by Nile Red staining. (B) Lipid metabolism-related proteins, PPARα and PGC-1α, were reduced in PA-treated cells. (C) Transcription levels of PPARα and PGC-1α were decreased by PA treatment. (D) Western blot results show that preproinsulin levels increased, and IRS2 and GLUT2 expression decreased in PA-treated cells in a concentration-dependent manner. (E) Transcription levels of insulin elevated and of IRS2 and GLUT2 declined following PA treatment. The upper panel shows representative electrophorogram of the gene bands from RT-PCR and lower panels show the corresponding histograms from real-time qPCR in C and E. *p<0.05, **p<0.01, ***p<0.001 compared with the control group.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibody including anti-human p62 polyclonal antibody (PM045, MBL, Nagoya, Japan) diluted 1:500, and anti-human β-ACTIN monoclonal antibody (TA-09, ZSGB-Bio, Beijing, China) diluted 1:1000, anti-human LC3B polyclonal antibody (PM036, MBL) diluted 1:500, anti-human LAMP2 monoclonal antibody (sc-5571, Santa Cruz Biotechnology, Dallas, TX) diluted 1:200, anti-human PPARA monoclonal antibody (AM8452b, Abgent, Suzhou, China) diluted 1:500, anti-human PGC-1α polyclonal antibody (ab54481, Abcam, Cambridge, UK) diluted 1:500, anti-human insulin polyclonal antibody (sc-7838,Santa Cruz Biotechnology) diluted 1:200, anti-human GLUT2 polyclonal antibody (sc-9117,Santa Cruz Biotechnology) diluted 1:200, and anti-human IRS2 polyclonal antibody (#4502, CST, Danvers, MA, USA) diluted 1:200.

Techniques: Concentration Assay, Staining, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Biological Sciences

Article Title: Intracellular Insulin and Impaired Autophagy in a Zebrafish model and a Cell Model of Type 2 diabetes

doi: 10.7150/ijbs.19249

Figure Lengend Snippet: Primers for qPCR of zebrafish genes used in this study

Article Snippet: The membranes were incubated overnight at 4°C with primary antibody including anti-human p62 polyclonal antibody (PM045, MBL, Nagoya, Japan) diluted 1:500, and anti-human β-ACTIN monoclonal antibody (TA-09, ZSGB-Bio, Beijing, China) diluted 1:1000, anti-human LC3B polyclonal antibody (PM036, MBL) diluted 1:500, anti-human LAMP2 monoclonal antibody (sc-5571, Santa Cruz Biotechnology, Dallas, TX) diluted 1:200, anti-human PPARA monoclonal antibody (AM8452b, Abgent, Suzhou, China) diluted 1:500, anti-human PGC-1α polyclonal antibody (ab54481, Abcam, Cambridge, UK) diluted 1:500, anti-human insulin polyclonal antibody (sc-7838,Santa Cruz Biotechnology) diluted 1:200, anti-human GLUT2 polyclonal antibody (sc-9117,Santa Cruz Biotechnology) diluted 1:200, and anti-human IRS2 polyclonal antibody (#4502, CST, Danvers, MA, USA) diluted 1:200.

Techniques: Sequencing

Journal: International Journal of Biological Sciences

Article Title: Intracellular Insulin and Impaired Autophagy in a Zebrafish model and a Cell Model of Type 2 diabetes

doi: 10.7150/ijbs.19249

Figure Lengend Snippet: Primers for qPCR of human genes used in this study

Article Snippet: The membranes were incubated overnight at 4°C with primary antibody including anti-human p62 polyclonal antibody (PM045, MBL, Nagoya, Japan) diluted 1:500, and anti-human β-ACTIN monoclonal antibody (TA-09, ZSGB-Bio, Beijing, China) diluted 1:1000, anti-human LC3B polyclonal antibody (PM036, MBL) diluted 1:500, anti-human LAMP2 monoclonal antibody (sc-5571, Santa Cruz Biotechnology, Dallas, TX) diluted 1:200, anti-human PPARA monoclonal antibody (AM8452b, Abgent, Suzhou, China) diluted 1:500, anti-human PGC-1α polyclonal antibody (ab54481, Abcam, Cambridge, UK) diluted 1:500, anti-human insulin polyclonal antibody (sc-7838,Santa Cruz Biotechnology) diluted 1:200, anti-human GLUT2 polyclonal antibody (sc-9117,Santa Cruz Biotechnology) diluted 1:200, and anti-human IRS2 polyclonal antibody (#4502, CST, Danvers, MA, USA) diluted 1:200.

Techniques: Sequencing

Journal: Biomedicines

Article Title: Understanding the Role of GLUT2 in Dysglycemia Associated with Fanconi–Bickel Syndrome

doi: 10.3390/biomedicines10092114

Figure Lengend Snippet: Genetic analysis for case 1 and family. ( a ) Sanger sequencing of DNA of the patient showed a novel homozygous mutation of SLC2A2 (C. 613-7T>GIVS 5-7T>G) expected to effect a splice site between exons 5 and 6. ( b ) cDNA sequencing demonstrates that the splicing of exons 5 and 6 are unaffected by the intronic mutation. Parents are carriers of the mutation.

Article Snippet: Cells were incubated with either 10 μL of anti-human GLUT2 PE-conjugated mouse IgG2a antibody (R&D SYSTEMS, FAB14148) or with FMO only.

Techniques: Sequencing, Mutagenesis

Journal: Biomedicines

Article Title: Understanding the Role of GLUT2 in Dysglycemia Associated with Fanconi–Bickel Syndrome

doi: 10.3390/biomedicines10092114

Figure Lengend Snippet: Genetic analysis for case 2 and family. Sanger sequencing of DNA shows a homozygous mutation of SLC2A2 (c.1093C>T in exon 9, R365X) in the patient. Both parents are carriers of the mutation.

Article Snippet: Cells were incubated with either 10 μL of anti-human GLUT2 PE-conjugated mouse IgG2a antibody (R&D SYSTEMS, FAB14148) or with FMO only.

Techniques: Sequencing, Mutagenesis

Journal: Biomedicines

Article Title: Understanding the Role of GLUT2 in Dysglycemia Associated with Fanconi–Bickel Syndrome

doi: 10.3390/biomedicines10092114

Figure Lengend Snippet: Flow cytometry to assess GLUT2 expression in each cell type in healthy PBMCs. ( a ) GLUT2 expression was very low in all cell populations in PBMCs extracted from the healthy control. ( b ) However, the expression of GLUT2 was increased in specific cell populations in PBMCs extracted from an immune-activated control.

Article Snippet: Cells were incubated with either 10 μL of anti-human GLUT2 PE-conjugated mouse IgG2a antibody (R&D SYSTEMS, FAB14148) or with FMO only.

Techniques: Flow Cytometry, Expressing, Control

Journal: Biomedicines

Article Title: Understanding the Role of GLUT2 in Dysglycemia Associated with Fanconi–Bickel Syndrome

doi: 10.3390/biomedicines10092114

Figure Lengend Snippet: Western blotting to assess the expression of GLUT2 in patient and control PBMCs. ( a ) The expression of GLUT2 was increased in the patient with the exonic mutation. ( b ) The expression of GLUT2 was similar in the patient with the intronic mutation in comparison to control. A smaller band was detected by the GLUT2 antibody in the sample obtained from the patient carrying the exonic mutation, indicated by the arrow. The quantification (fold change) of GLUT2 in the samples are presented to the right in both ( a , b ).

Article Snippet: Cells were incubated with either 10 μL of anti-human GLUT2 PE-conjugated mouse IgG2a antibody (R&D SYSTEMS, FAB14148) or with FMO only.

Techniques: Western Blot, Expressing, Control, Mutagenesis, Comparison

Journal: Biomedicines

Article Title: Understanding the Role of GLUT2 in Dysglycemia Associated with Fanconi–Bickel Syndrome

doi: 10.3390/biomedicines10092114

Figure Lengend Snippet: qRT-PCR to assess the expression of glucose transporters in patient PBMCs. ( a ) The mRNA levels of both GLUT1 and GLUT3 are elevated in PBMCs obtained from the patient carrying the exonic mutation. ( b ) In contrast, the mRNA levels of GLUT1 and GLUT3 are reduced in PBMCs obtained from the patient carrying the intronic GLUT2 mutation. Error bar is the average of technical replicates.

Article Snippet: Cells were incubated with either 10 μL of anti-human GLUT2 PE-conjugated mouse IgG2a antibody (R&D SYSTEMS, FAB14148) or with FMO only.

Techniques: Quantitative RT-PCR, Expressing, Mutagenesis