Journal: Stem Cell Research & Therapy

Article Title: Insulin-like growth factor-II overexpression accelerates parthenogenetic stem cell differentiation into cardiomyocytes and improves cardiac function after acute myocardial infarction in mice

doi: 10.1186/s13287-020-1575-4

Figure Lengend Snippet: IGF-II overexpression promotes cardiomyocyte maturation in vivo. ESC- and PSC-derived cardiomyocytes at 1 × 10 5 cells/recipient mouse were injected into the heart at 1 week after acute myocardial infarction (MI). The IGF-II and VEGF levels in the injection sites ( a ) and the serum ( b ) were measured by ELISA at 4 weeks post-transplant. Data are expressed as the mean ± SD. * p < 0.05 vs. any other group. n = 12. c Immunofluorescence staining was performed to observe the cardiomyocyte morphology and to detect α-actinin expression (red staining) in the injection sites at 4 weeks post-transplant. Blue staining, Hoechst 33342. Green staining, eGFP. Scale bars, 10 μm

Article Snippet: IGF-II levels in serum (Quantikine ELISA Kit, cat# MG200, R&D Systems, Minneapolis, MN, USA), heart tissue (DuoSet ELISA, cat# DY792, R&D Systems), vascular endothelial growth factor (VEGF) levels in serum (cat# ab100751; Abcam), and heart tissue (cat# ab209882; Abcam) were determined with ELISA kits following the manufacturer’s instructions.

Techniques: Over Expression, In Vivo, Derivative Assay, Injection, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Expressing

Journal: bioRxiv

Article Title: Hematopoietic Stem and Progenitor Cell Aging is Initiated at Middle Age Through Decline in Local Insulin-Like Growth Factor 1 (IGF1)

doi: 10.1101/2020.07.11.198846

Figure Lengend Snippet: a ) Expression pattern of Igf2 , Nrg1 , Tgfb1 , and Egf in BM subsets assessed by scRNA-seq . For detailed cell type annotation refer to: https://nicheview.shiny.embl.de . ( b ) IGF2 concentration in BM fluid of a single mouse femur from 2-4mo ( n = 7), 6mo (n = 6), 12-14mo ( n = 5) and 26-28mo ( n = 7) mice. ( c ) Expression of Igf1r in BM cells by scRNA-seq . ( d ) Left: Experimental design. Right: Frequency of donor cells (CD45.1+) and donor-derived T cells (CD45.1+ CD3+) in the blood at 24wks post-transplant of WT BM into lethally irradiated WT or Igf1 fl/fl ; CreER T2 recipient mice ( n = 5, 7, from left to right). ( e ) Left: Experimental design. Right: Frequency of donor cells (CD45.2+), and donor-derived myeloid cells (CD11b+), B cells (B220+), and T cells (CD3+) in the blood at 24wks post-transplant of WT or Igf1r fl/fl Mx1 -Cre BM into lethally irradiated WT recipient mice ( n = 7, 4, from left to right). ( f ) Left: Experimental design. Right: Frequency of donor cells (CD45.1+) and donor-derived T cells (CD45.1+ CD3+) in the blood at 20wks post-transplant of WT BM into lethally irradiated WT or Igf1 fl/fl ; Nestin-Cre ER recipient mice ( n = 6, 5, from left to right). ( g ) Top panel: Igf1 recombination PCR, middle panel: Igf1 genotyping PCR and bottom panel: Cre genotyping PCR. Input for all reactions was gDNA isolated from livers of tamoxifen-treated Igf1 +/+ or Igf1 fl/fl ; Nestin-Cre ER mice or recombined control (CNT; Igf1 fl/fl ; Cre-ER T2 ) ( n = 3, 3, from left to right). ( b, d-f ) Dots represent individual mice and bars are mean ± SEM. P -values were generated for ( b ) by one-way ANOVA with Holm-Sidak’s multiple comparisons test, ( d - f ) by unpaired, two-tailed t tests.

Article Snippet: A mouse-specific IGF1 Immunoassay kit (ELISA, R&D Systems) and a mouse-specific IGF2 PicoKine kit (ELISA, Boster biological technology) was used to determine the concentration of IGF1 and IGF2 proteins in the BM.

Techniques: Expressing, Concentration Assay, Derivative Assay, Irradiation, Isolation, Control, Generated, Two Tailed Test

Journal: Communications Biology

Article Title: Paradoxical regulation of IGF2 in promoting lipid metabolism in adipose tissues

doi: 10.1038/s42003-025-08458-1

Figure Lengend Snippet: A , B “U” and inverted “U” shaped relationship between IGF2 levels and the lipid species detected by LC-MS/MS assay utilizing a polynomial fourth order equation to fit the non-linear regression curve, where R² (R-squared) represents the coefficient of determination, and Sy.x represents the standard deviation of the residuals, n = 200. C , D “U” and inverted “U” shaped relationship between IGF2 levels and the lipid species detected by ELISA kit assay utilizing a polynomial fourth order equation to fit the non-linear regression curve, n = 200. E , F Pearson correlation analyses of L-IGF2 levels and H-IGF2 levels with triglyceride conducted by LC-MS/MS assay, n = 200, all p < 0.01. G , H Pearson correlation analyses of L-IGF2 levels and H-IGF2 levels with HDL-c conducted by LC-MS/MS assay, n = 200, all p < 0.05. I , J Pearson correlation analyses of L-IGF2 levels and H-IGF2 levels with triglyceride conducted by ELISA kit assay, n = 200, all p < 0.001. K , L Pearson correlation analyses of L-IGF2 levels and H-IGF2 levels with HDL-c conducted by ELISA kit assay, n = 200, all p < 0.05. M , N Multiple stepwise logistic regression analysis of MetS, HOMA-IR, and other metabolic subgroups (central obesity, hypertension, hyperglycemia, hypertriglyceridemia, and low HDL-c) connected with L-IGF2 and H-IGF2 levels conducted by LC-MS/MS assay and ELISA kit assay, respectively.

Article Snippet: The measurement of serum IGF2 level was conducted with an ELISA kit (E-EL-H6037 for human samples and E-EL-M3078 for mice samples, Elabscience, China).

Techniques: Liquid Chromatography with Mass Spectroscopy, Standard Deviation, Enzyme-linked Immunosorbent Assay

Journal: Communications Biology

Article Title: Paradoxical regulation of IGF2 in promoting lipid metabolism in adipose tissues

doi: 10.1038/s42003-025-08458-1

Figure Lengend Snippet: A The diagram of Ad-IGF2 and its control adenovirus were injected into the eWAT of DIO mice at multi-point (2 × 10 10 pfu/mouse, injected two times for a week, a total of 4 weeks of injection). B In vivo imaging was employed to track the expression sites of the overexpressing adenovirus constructs tagged with GFP, including Ad-IGF2 and Ad-GFP. C , D Representative western blot results of IGF2 protein levels in iWAT, eWAT, liver, skeletal muscle and pancreas tissues. E For a duration of 8 weeks, eWAT and iWAT of the mice were carefully dissected and visually documented using photography. F ELISA results of IGF2 protein levels in the serum. n = 6, ** p < 0.01 by two-tailed, unpaired Student’s t test. G – J The weight of eWAT and iWAT, liver and the whole-body weight of mice were compared, n = 6, * p < 0.05, n.s not significant (unpaired Student’s t test). K Hematoxylin/Eosin representative staining of adipocytes in iWAT, eWAT, prWAT, liver, and muscle tissue. Magnification: 20×. L – N Adipocyte size distribution curves of eWAT, iWAT and prWAT between Ad-GFP group mice (gray) and Ad-IGF2 group mice (blue), respectively. n = 6, Data represent the mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001 analyzed by a one-way ANOVA with a Tukey’s multiple comparisons test. O , P The measurement analysis of blood glucose levels was conducted at specific times in obese mice after IGF2 overexpression by GTT and ITT assays, n = 6, * p < 0.05, analyzed by a two-way ANOVA with Bonferroni’s multiple comparisons test. Q , R The area under curve (AUC) analysis of GTT and ITT assays were conducted in obese mice after IGF2 overexpression, n = 6, *** p < 0.001 by two-tailed, unpaired Student’s t test. S , T Plasma TC, TG, HDL-c, LDL-c, ALT, AST, and ALP contents have been identified following injection of Ad-GFP and Ad-IGF2, n = 6, Data represent the mean ± SD; * p < 0.05 (unpaired Student’s t test).

Article Snippet: The measurement of serum IGF2 level was conducted with an ELISA kit (E-EL-H6037 for human samples and E-EL-M3078 for mice samples, Elabscience, China).

Techniques: Control, Injection, In Vivo Imaging, Expressing, Construct, Western Blot, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Staining, Over Expression, Clinical Proteomics

Journal: Communications Biology

Article Title: Paradoxical regulation of IGF2 in promoting lipid metabolism in adipose tissues

doi: 10.1038/s42003-025-08458-1

Figure Lengend Snippet: A The diagram of IGF2-RNAi and its control lentivirus NC-RNAi were injected into the tail vein of 3-4-week-old mice (1 × 10 10 pfu/mouse, injected two times for a week, collectively a 4 weeks of injection). B Representative western blot results of IGF2 protein levels in liver tissues when 16 weeks old. C Representative western blot results of IGF2 protein levels in eWAT, iWAT, liver, skeletal muscle, pancreas and kidney tissues. D ELISA results of IGF2 protein level in serum samples, n = 12, *** p < 0.001 (unpaired Student’s t test). E At 16 weeks, liver tissues, iWAT, and eWAT of mice underwent dissection and photography. The representative images were depicted. F – J The whole-body weight, the liver, iWAT, eWAT ( n = 12) and quadriceps ( n = 6) weight of mice were compared, the p -values have been annotated, n.s not significant (unpaired Student’s t test). K Representative H&E staining of iWAT, eWAT, liver and muscle tissues, and the Oil Red staining of liver and muscle tissues. Magnification: 20×. L , M Adipocyte size distribution curves of eWAT and iWAT between NC-RNAi group mice (gray) and IGF2-RNAi group mice (yellow), respectively. n = 6, * p < 0.05, ** p < 0.01, *** p < 0.001 analyzed by a one-way ANOVA with a Tukey’s multiple comparisons test. N , O Plasma TC, TG, HDL-c, LDL-c, ALT, AST, and ALP contents have been identified following injection of IGF2-RNAi and NC-RNAi, n = 6, * p < 0.05, ** p < 0.01 (unpaired Student’s t test). P , R The levels of blood glucose underwent measurement at specific times by GTT and ITT assays, n = 6, * p < 0.05, ** p < 0.01 analyzed by a two-way ANOVA with Bonferroni’s multiple comparisons test. Q , S The area under curve (AUC) analysis of GTT and ITT assays were conducted in obese mice after IGF2 knockdown, n = 6, Data represent the mean ± SD; *** p < 0.001 by two-tailed, unpaired Student’s t test.

Article Snippet: The measurement of serum IGF2 level was conducted with an ELISA kit (E-EL-H6037 for human samples and E-EL-M3078 for mice samples, Elabscience, China).

Techniques: Control, Injection, Western Blot, Enzyme-linked Immunosorbent Assay, Dissection, Staining, Clinical Proteomics, Knockdown, Two Tailed Test

Journal: Communications Biology

Article Title: Paradoxical regulation of IGF2 in promoting lipid metabolism in adipose tissues

doi: 10.1038/s42003-025-08458-1

Figure Lengend Snippet: A The utilization of Nile red staining and Oil red O staining techniques revealed the presence of lipid droplets in 3T3-L1 adipocytes after IGF2 overexpression and knockdown treatment. B , C Quantification of cellular triglyceride (TG) content and the levels of free fatty acid (FFA) in the medium in 3T3-L1 adipocytes after IGF2 overexpression and knockdown treatment. * p < 0.05, ** p < 0.01. D , E The mRNA levels of genes associated with adipogenesis, lipogenesis, and lipolysis in 3T3-L1 cells transfected with Ad-IGF2 or IGF2-RNAi as well as the corresponding controls by RT-qPCR assays, using Ppia as internal controls, n = 6, Data represent the mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001 by two-tailed, unpaired Student’s t test. F Protein expression levels of genes related to adipogenesis, lipogenesis and lipolysis in the 3T3-L1 adipocytes by western blot assays. G Mice adipose tissue has been dissected and isolated for primary adipocyte culture, and the results of 0, 4, 8 and 12 days were induced by the classic “Cocktail” induction differentiation regimen. H Primal cell supernatant was extracted during the above induction differentiation process, and IGF2 concentration was detected by ELISA kit. n = 3. I 3T3-L1 preadipocytes underwent treatment via different concentration gradients of recombinant IGF2 protein powder and induced differentiation. Oil red O staining employment demonstrated that the deposition of lipid droplets within adipocytes.

Article Snippet: The measurement of serum IGF2 level was conducted with an ELISA kit (E-EL-H6037 for human samples and E-EL-M3078 for mice samples, Elabscience, China).

Techniques: Staining, Over Expression, Knockdown, Transfection, Quantitative RT-PCR, Two Tailed Test, Expressing, Western Blot, Isolation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Recombinant