recombinant human igf2  (Sino Biological)

Journal: iScience

Article Title: Reprogramming of fibroblasts into cancer-associated fibroblasts via IGF2-mediated autophagy promotes metastasis of lung cancer cells

doi: 10.1016/j.isci.2024.111269

Figure Lengend Snippet: Lung cancer cells induced CAFs activation via IGF2 secretion (A) mRNA levels of IGF2 in cells were detected by qPCR. (B) IGF2 concentrations in cell culture medium were detected by ELISA. (C) NFs were treated with IGF2 (50 and 100 ng/mL) for 24 h. Cell morphology was observed under a microscope. (D) NFs were treated with IGF2. α-SMA and FAP expressions were detected by western blotting. (E) NFs were treated with IGF2. Cell proliferation was examined by CCK-8 kit after 48 h. (F and G) NFs were treated with IGF2. NFs migration was detected by trans-well assay after 24 h. (H) NFs were treated with CM or CM + IGF2 neutralizing antibody. Cell morphology was observed under a microscope. (I) NFs were treated with CM or CM + IGF2 neutralizing antibody. α-SMA and FAP expressions were detected by western blotting. (J) NFs were treated with CM or CM + IGF2 neutralizing antibody, and IGF2 (100 ng/mL). Cell proliferation was examined by CCK-8 kit after 48 h. (K and L) NFs were treated with CM or CM + IGF2 neutralizing antibody. NFs migration was detected by trans-well assay after 24 h. (M) The effect of IGF2 knockdown by RNAi. (N) IGF2 expression was knocked down in lung cancer cells, and then the CM was collected and used to culture NFs. α-SMA and FAP expressions were detected by western blotting. (O and P) IGF2 expression was knocked down in lung cancer cells by RNAi, and then the CM was collected and used to culture NFs. NFs migration was detected by trans-well assay after 24 h. Ab, neutralizing antibody. Scale bar, 100 μm. Data represents the mean ± SD from three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 .

Article Snippet: Recombinant human IGF2 , Sino Biological , Cat#13032-HNAY.

Techniques: Activation Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Microscopy, Western Blot, CCK-8 Assay, Migration, Knockdown, Expressing

Journal: iScience

Article Title: Reprogramming of fibroblasts into cancer-associated fibroblasts via IGF2-mediated autophagy promotes metastasis of lung cancer cells

doi: 10.1016/j.isci.2024.111269

Figure Lengend Snippet: IGF2-induced autophagy mediated the effect of lung cancer cells on CAFs activation (A) NFs were treated with CM and then were stained with 0.5 μg/mL AO for 15 min. The acidic vesicular organelles (AVOs) formation was observed under a fluorescence microscope (scale bar, 20 μm). (B) NFs were treated with CM. The expression of LC-3II and ATG5 were detected by western blotting. (C) NFs were treated with IGF2 (50 or 100 mg/mL) for 24 h. The AVOs formation was examined by AO staining (scale bar, 20 μm). (D) NFs were treated with IGF2. The expression of LC-3II and ATG5 was detected by western blotting. (E) NFs were treated with IGF2 or IGF2 + 3-MA (5 mM). The AVOs formation was examined by AO staining (scale bar, 20 μm). (F) NFs were treated with IGF2 or IGF2 + 3-MA. Protein expressions were detected by western blotting. (G) NFs were treated with IGF2 or IGF2 + 3-MA. Cell proliferation was examined by CCK-8 kit after 48 h. (H and I) NFs were treated with IGF2 or IGF2+3-MA. NFs migration was detected by trans-well assay after 24 h (scale bar, 100 μm). (J) NFs were treated with IGF2, CM, CM + IGF2 Ab, and CM + 3-MA. The AVOs formation was examined by AO staining (scale bar, 20 μm). (K) NFs were treated with CM, IGF2, and CM + IGF2 Ab. Protein expressions were detected by western blotting. (L) NFs were treated with IGF2, CM, CM + IGF2 Ab, and CM + 3-MA. Cell proliferation was examined by CCK-8 kit after 48 h. (M and N) NFs were treated with IGF2, CM, CM + IGF2 Ab, and CM + 3-MA. NFs migration was detected by trans-well assay after 24 h (scale bar, 100 μm). (O and P) NFs were treated with RAPA (100 nM); α-SMA and FAP expressions were detected by western blotting (scale bar, 20 μm). (Q and R) NFs were treated with RAPA; NFs migration was detected by trans-well assay after 24 h. Ab, neutralizing antibody (scale bar, 100 μm). Data represents the mean ± SD from three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 .

Article Snippet: Recombinant human IGF2 , Sino Biological , Cat#13032-HNAY.

Techniques: Activation Assay, Staining, Fluorescence, Microscopy, Expressing, Western Blot, CCK-8 Assay, Migration

Journal: iScience

Article Title: Reprogramming of fibroblasts into cancer-associated fibroblasts via IGF2-mediated autophagy promotes metastasis of lung cancer cells

doi: 10.1016/j.isci.2024.111269

Figure Lengend Snippet:

Article Snippet: Recombinant human IGF2 , Sino Biological , Cat#13032-HNAY.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Reprogramming of fibroblasts into cancer-associated fibroblasts via IGF2-mediated autophagy promotes metastasis of lung cancer cells

doi: 10.1016/j.isci.2024.111269

Figure Lengend Snippet: PCR primer sequences

Article Snippet: Recombinant human IGF2 , Sino Biological , Cat#13032-HNAY.

Techniques:

Journal: Frontiers in Immunology

Article Title: PTN/IGF-2 signaling modulates endometrial decidualization and immune cell trafficking to facilitate pregnancy maintenance

doi: 10.3389/fimmu.2026.1790942

Figure Lengend Snippet: PTN regulates endometrial stromal cell decidualization through the IGF-2 signaling axis. (A) PPI network of PTN, IGF-2, IGFBP1, LIF, PRL, and other decidualization-related genes. (B) The expression levels of PTN and its interacting partners (IGF-2, WNT4, PRL, IGFBP1, and LIF) in the RIF (left) and RPL (right) datasets. (C) Expressions of decidualization-related genes (including IGFBP1 , LIF , PRL , and WNT4 ) in NC or si PTN hESCs after treatment with control vehicle, cAMP, and IGF-2 via RT-qPCR (n = 9). (D) Immunoblotting for PTN, IGFBP1, and PRL expression levels with control vehicle, cAMP, and IGF-2. (E) ELISA for IGF-2 levels with control vehicle, cAMP, and IGF-2. (F) Schematic diagram of the experiments performed using the different treatments of hESCs. Data are presented as mean ± SEM and analyzed using t-test or one-way ANOVA test. * compared with NC treated with control vehicle, # compared with si PTN treated with control vehicle, $ compared with si PTN treated with control cAMP (NS, no significant difference; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ####p < 0.0001, $p < 0.05, $$p < 0.01, $$$p < 0.001, $$$$p< 0.0001). PPI, protein–protein interaction; RIF, recurrent implantation failure; RPL, recurrent pregnancy loss; hESCs, human endometrial stromal cells; cAMP, cyclic adenosine monophosphate.

Article Snippet: The remaining cells were digested with trypsin and inoculated into a new culture dish, and they were treated with control vehicle or IGF-2 (50 ng/mL, R&D Systems, Minneapolis, MN, USA, 292-G2) for 48 h and then collected for quantitative real-time polymerase chain reaction (qRT-PCR).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: PTN/IGF-2 signaling modulates endometrial decidualization and immune cell trafficking to facilitate pregnancy maintenance

doi: 10.3389/fimmu.2026.1790942

Figure Lengend Snippet: IGF-2 promotes decidualization and rescues decidualization defects caused by PTN deficiency. (A) Schematic diagram of the experiments performed using the different treatments of ESCs isolated from endometrium of RIF and RPL patients. (B, C) RT-qPCR analysis of levels of decidualization-related genes in ESCs isolated from endometrium of RIF (B) and RPL (C) patients after IGF-2 (50 ng/mL) treatment for 48 h (n = 9). (D) The flowchart depicts the steps involved in establishing a mouse model of intrauterine perfusion with siRNA-mediated knockdown of PTN and then supplemented with IGF-2 (50 ng/mL) through tail vein injection. (E) Expression of PTN after Ptn knockdown and supplemented with control vehicle or IGF-2 (50 ng/mL) in the mouse endometrium, measured by immunofluorescence, and quantitative analysis of immunofluorescence was performed (n = 6). (F) RT-qPCR analysis of levels of decidualization-related genes after Ptn knockdown and supplemented with control vehicle or IGF-2 (50 ng/mL) in the mouse endometrium (n = 6). Data are presented as mean ± SEM and analyzed using t-test or one-way ANOVA test. * compared with NC treated with control vehicle, # compared with si PTN treated with control vehicle (NS, no significant difference; *p < 0.05, **p < 0.01, ***p < 0.001, ****p< 0.0001, ## p < 0.01, ### p < 0.001, #### p < 0.0001). ESCs, endometrial stromal cells; PPI, protein–protein interaction; RIF, recurrent implantation failure.

Article Snippet: The remaining cells were digested with trypsin and inoculated into a new culture dish, and they were treated with control vehicle or IGF-2 (50 ng/mL, R&D Systems, Minneapolis, MN, USA, 292-G2) for 48 h and then collected for quantitative real-time polymerase chain reaction (qRT-PCR).

Techniques: Isolation, Quantitative RT-PCR, Knockdown, Injection, Expressing, Control, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: PTN/IGF-2 signaling modulates endometrial decidualization and immune cell trafficking to facilitate pregnancy maintenance

doi: 10.3389/fimmu.2026.1790942

Figure Lengend Snippet: PTN deficiency induces endometrial immune imbalance and leads to adverse pregnancy outcomes, which can be partially reversed by IGF-2. (A) PPI network of PTN, IGF-2, CXCR4, CD4, CD8, CD56, and other immunoregulators. (B, C) Flow cytometry analysis (B) and statistical quantification (C) of cell populations of NK cells and T cells, and the expression levels of CD16, CXCR4, and GZMB in NK cell form the endometrial tissues of NC and si PTN mice treated with control vehicle or IGF-2 (50 ng/mL) (n = 6). (D) The flowchart depicts the steps involved in establishing a mouse model of intrauterine perfusion with siRNA-mediated knockdown of Ptn and then supplemented with IGF-2 (50 ng/mL) through tail vein injection for 2 days; then, these female mice were mated with fertile male mice; analysis of fertility, including pregnancy rate, IF, and fluorescence-activated cell sorting (FACS). were performed at gestational day 13.5. (E) Representative images of uteri from pregnant mice in the NC and si PTN mice treated with control vehicle or IGF-2 (50 ng/mL) at gestational day 13.5 (n = 6) (arrow shows the absorption site). (F) Pregnancy rate (%) in NC and si PTN mice treated with control vehicle or IGF-2 (50 ng/mL) (n = 15). (G) Quantification of embryo numbers (left) and absorption rate (right) per mouse in NC (n = 13) and si PTN mice treated with control vehicle (n = 6) or IGF-2 (50 ng/mL) (n = 11) at gestational day 13.5. Data are presented as mean ± SEM and analyzed using one-way ANOVA test or χ 2 test. * Compared with NC treated with control vehicle, # compared with si PTN treated with control vehicle (NS, no significant difference; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ## p < 0.01, ### p < 0.001, #### p < 0.0001). IF, immunofluorescence.

Article Snippet: The remaining cells were digested with trypsin and inoculated into a new culture dish, and they were treated with control vehicle or IGF-2 (50 ng/mL, R&D Systems, Minneapolis, MN, USA, 292-G2) for 48 h and then collected for quantitative real-time polymerase chain reaction (qRT-PCR).

Techniques: Flow Cytometry, Expressing, Control, Knockdown, Injection, Fluorescence, FACS, Immunofluorescence

Journal: The Journal of Reproduction and Development

Article Title: The imprinted Igf2-Igf2r axis is critical for exosome biogenesis during the early development of bovine placenta

doi: 10.1262/jrd.2024-081

Figure Lengend Snippet: Histological analysis of cotyledon villi of bovine placenta at 2, 3, and 4 months of gestation. (A) The cotyledon villi at 2, 3, and 4 months of pregnancy were collected for HE staining; the black arrows indicate mononuclear trophoblasts (MNCs) and binuclear trophoblasts (BNCs), Scale bars = 100 μm. Plasma was collected from pregnant dairy cows at 60, 90, and 120 days gestation, and the secretion of IGF2 (B) and IGF2R (C) was detected using ELISA kits according to the manufacturer’s recommendations. Data represent the mean ± standard deviations from six independent experiments. Bars with different letters indicate significant differences (* P < 0.05).

Article Snippet: After fixation, the tissues were washed three times with PBS and permeabilized with PBS containing 0.2% (v/v) Triton X-100 (Applygen, Beijing, China), and 50 μl of TSG101 (1:200 dilution, bs-1365R, Bioss), Rab11 (1:200 dilution, 15903-1-AP, Proteintech), CD63 (1:200 dilution; bs-1523R; Bioss), CD9 (1:200 dilution; bs-2486R; Bioss), IGF2R (1:200 dilution; 14364S; Cell Signaling Technology) and IGF2 (1:200 dilution; 12220-1-AP; Proteintech) primary antibody, lightly washed five times with PBS, and incubated with 1:100 diluted fluorescent secondary antibody (1:200 dilution; TransGen Biotech) for 1 h at room temperature in the dark.

Techniques: Staining, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Reproduction and Development

Article Title: The imprinted Igf2-Igf2r axis is critical for exosome biogenesis during the early development of bovine placenta

doi: 10.1262/jrd.2024-081

Figure Lengend Snippet: Localization and expression of IGF2, IGF2R, and exosomal marker proteins. (A) Immunohistochemical assays were utilized to detect the localization of IGF2, IGF2R, and exosomal marker proteins CD9, CD63, TSG101, and Rab11 in the cotyledon villi of bovine placenta at 2, 3, and 4 months of gestation. Scale bar = 50 μm. (B) Western blot analysis was utilized to detect the expressions of IGF2, IGF2R, and exosomal marker proteins CD63, CD9, TSG101, and Rab11 in the cotyledon villi of bovine placenta at 2, 3, and 4 months of gestation. (C) Band intensity measurement of the IGF2, IGF2R, and exosomal marker proteins CD63, CD9, TSG101, and Rab11 to β-actin were determined by densitometry. The data represent the mean ± standard deviations from three independent experiments. Bars with different letters indicate statistically significant differences (* P < 0.05).

Article Snippet: After fixation, the tissues were washed three times with PBS and permeabilized with PBS containing 0.2% (v/v) Triton X-100 (Applygen, Beijing, China), and 50 μl of TSG101 (1:200 dilution, bs-1365R, Bioss), Rab11 (1:200 dilution, 15903-1-AP, Proteintech), CD63 (1:200 dilution; bs-1523R; Bioss), CD9 (1:200 dilution; bs-2486R; Bioss), IGF2R (1:200 dilution; 14364S; Cell Signaling Technology) and IGF2 (1:200 dilution; 12220-1-AP; Proteintech) primary antibody, lightly washed five times with PBS, and incubated with 1:100 diluted fluorescent secondary antibody (1:200 dilution; TransGen Biotech) for 1 h at room temperature in the dark.

Techniques: Expressing, Marker, Immunohistochemical staining, Western Blot

Journal: The Journal of Reproduction and Development

Article Title: The imprinted Igf2-Igf2r axis is critical for exosome biogenesis during the early development of bovine placenta

doi: 10.1262/jrd.2024-081

Figure Lengend Snippet: Knockdown of Igf2r gene in BTCs represses the expression of exosome biogenesis-related proteins in BTCs and BTCs-derived exosomes. (A) The expressions of IGF2, IGF2R, CD63, CD9, TSG101, and Rab11 proteins in Igf2r knockdown BTCs were detected using western blot analysis. (B) Quantification of band intensities was determined using densitometric analysis. The data represent the mean ± standard deviations from three independent experiments. Bars with different letters indicate statistically significant differences (* P < 0.05). (C) The expressions of CD63, CD9, TSG101, and Rab11 proteins in Igf2r knockdown BTCs-derived exosomes were identified with western blot analysis. (D) Quantification of band intensities was determined using densitometric analysis. The data represent the mean ± standard deviations from three independent experiments. Bars with different letters indicate statistically significant differences (* P < 0.05).

Article Snippet: After fixation, the tissues were washed three times with PBS and permeabilized with PBS containing 0.2% (v/v) Triton X-100 (Applygen, Beijing, China), and 50 μl of TSG101 (1:200 dilution, bs-1365R, Bioss), Rab11 (1:200 dilution, 15903-1-AP, Proteintech), CD63 (1:200 dilution; bs-1523R; Bioss), CD9 (1:200 dilution; bs-2486R; Bioss), IGF2R (1:200 dilution; 14364S; Cell Signaling Technology) and IGF2 (1:200 dilution; 12220-1-AP; Proteintech) primary antibody, lightly washed five times with PBS, and incubated with 1:100 diluted fluorescent secondary antibody (1:200 dilution; TransGen Biotech) for 1 h at room temperature in the dark.

Techniques: Knockdown, Expressing, Derivative Assay, Western Blot

Journal: The Journal of Reproduction and Development

Article Title: The imprinted Igf2-Igf2r axis is critical for exosome biogenesis during the early development of bovine placenta

doi: 10.1262/jrd.2024-081

Figure Lengend Snippet: Repression of IGF2 protein expression inhibits the expression of exosomal marker proteins in BTCs and BTC-derived exosomes. (A) The expressions of IGF2, IGF2R, CD63, CD9, TSG101, and Rab11 proteins in BTCs treated with chromeceptin for 24 h were detected with western blot analysis. (B) Quantification of band intensities was determined using densitometric analysis. The data represent the mean ± standard deviations from three independent experiments. Bars with different letters indicate statistically significant differences (* P < 0.05). (C) The expressions of CD63, CD9, TSG101, and Rab11 proteins in exosomes derived from BTCs treated with chromeceptin were detected with western blot analysis. (D) Quantification of band intensities was determined using densitometric analysis. The data represent the mean ± standard deviations from three independent experiments. Bars with different letters indicate statistically significant differences (* P < 0.05).

Article Snippet: After fixation, the tissues were washed three times with PBS and permeabilized with PBS containing 0.2% (v/v) Triton X-100 (Applygen, Beijing, China), and 50 μl of TSG101 (1:200 dilution, bs-1365R, Bioss), Rab11 (1:200 dilution, 15903-1-AP, Proteintech), CD63 (1:200 dilution; bs-1523R; Bioss), CD9 (1:200 dilution; bs-2486R; Bioss), IGF2R (1:200 dilution; 14364S; Cell Signaling Technology) and IGF2 (1:200 dilution; 12220-1-AP; Proteintech) primary antibody, lightly washed five times with PBS, and incubated with 1:100 diluted fluorescent secondary antibody (1:200 dilution; TransGen Biotech) for 1 h at room temperature in the dark.

Techniques: Expressing, Marker, Derivative Assay, Western Blot