Journal: Molecular and Cellular Biology

Article Title: Human Insulin Growth Factor 2 mRNA Binding Protein 2 Increases MicroRNA 33a/b Inhibition of Liver ABCA1 Expression and Alters Low-Density Apolipoprotein Levels in Mice

doi: 10.1128/MCB.00058-20

Figure Lengend Snippet: IGF2BP2 is required for miR-33a/b-dependent silencing of ABCA1 mRNA expression. (A) Luciferase activity was quantitated in HepG2 cells transfected with a control luciferase reporter (Con-Luc), ABCA1 3′-UTR-containing reporter (3′ UTR), ABCA1 3′-UTR mutant reporter (MUT-3′ UTR), pre-miR-33a/b (miR-33a/b), or control pre-miR (Con-miR). Luciferase activity was measured as described in Materials and Methods. (B) Luciferase reporter activity was measured in cells treated with IGF2BP2 siRNA (IGF2BPsi) or transfected with pCMV-IGF2BP2 (IGF2BP2 OE). (C) qRT-PCR analysis was used to determine relative endogenous ABCA1 mRNA expression in miR-33a/b-expressing cells treated with IGF2BP2 siRNA or transfected with pCMV-IGF2BP2. ABCA1 mRNA expression values are compared to GAPDH mRNA expression, which was set at 1. (D) ABCA1 protein level was determined in miR-33a/b-expressing cells treated with IGF2BP2 siRNA or transfected with pCMV-IGF2BP2. Actin was used as a loading control. (E) qRT-PCR analysis was used to determine relative endogenous ABCA1 mRNA expression in miR-33a/b-expressing cells transfected with a miR-33a/b anti-miR, treated with IGF2BP2 siRNA, or transfected with pCMV-IGF2BP2. ABCA1 mRNA expression values were compared to GAPDH mRNA expression, which was set at 1. (F) ABCA1 protein level in miR-33a/b-expressing cells were transfected with a miR-33a/b anti-miR, treated with IGF2BP2 siRNA, or transfected with pCMV-IGF2BP2. Actin was used as a loading control. (G) Relative miR-33a/b mRNA abundance. (H) Relative ABCA1 mRNA abundance. **, P < 0.001; ***, P < 0.0001. The qRT-PCR data are the averages from five independent experiments. Values are the means ± SEM. The Western blot panels represent the average results of five independent experiments.

Article Snippet: Anti-ABCA1 polyclonal antibodies were from Abcam (ab1880) and used at a 1:500 dilution; anti-IGF2BP1 monoclonal antibodies were from Santa Cruz (IGF2BP1 [sc-390149], IGF2BP2 [sc-377014], and IGF2BP3 [sc-365640]), and all were used at a 1:2,000 dilution; anti-AGO2 polyclonal antibodies were from Abcam (ab32381) and used at a 1:750 dilution; anti-STAU1 polyclonal antibodies were from Sigma-Aldrich (SAB1406490) and used at a 1:1,000 dilution; anti-G3BP polyclonal antibodies were from Thermo Fisher (G3BP1 [PA5-29455]; G3BP2 [PA5-53797]) and were used at a 1:500 dilution.

Techniques: Expressing, Luciferase, Activity Assay, Transfection, Control, Mutagenesis, Quantitative RT-PCR, Western Blot

Journal: Molecular and Cellular Biology

Article Title: Human Insulin Growth Factor 2 mRNA Binding Protein 2 Increases MicroRNA 33a/b Inhibition of Liver ABCA1 Expression and Alters Low-Density Apolipoprotein Levels in Mice

doi: 10.1128/MCB.00058-20

Figure Lengend Snippet: IGF2BP2 binds the RISC endonuclease AGO2. miR-33a/b was biotinylated and incubated with cell lysates. Bound proteins were pulled down using streptavidin beads, resolved by SDS-PAGE, and analyzed by Western blotting. (A) Ten percent of the total protein level in the cell lysates is shown. (B) miR-33a/b was biotinylated and incubated with cell lysates. Bound proteins were pulled down using streptavidin beads, resolved by SDS-PAGE, and analyzed by Western blotting. Lysate, total cell lysate; scrambled, nonspecific miR control. (C) Cell lysates were incubated with polyclonal antibodies (IP; indicated at the top of the blots), and the levels of bound proteins were determined. The figure represents the average results of five independent experiments.

Article Snippet: Anti-ABCA1 polyclonal antibodies were from Abcam (ab1880) and used at a 1:500 dilution; anti-IGF2BP1 monoclonal antibodies were from Santa Cruz (IGF2BP1 [sc-390149], IGF2BP2 [sc-377014], and IGF2BP3 [sc-365640]), and all were used at a 1:2,000 dilution; anti-AGO2 polyclonal antibodies were from Abcam (ab32381) and used at a 1:750 dilution; anti-STAU1 polyclonal antibodies were from Sigma-Aldrich (SAB1406490) and used at a 1:1,000 dilution; anti-G3BP polyclonal antibodies were from Thermo Fisher (G3BP1 [PA5-29455]; G3BP2 [PA5-53797]) and were used at a 1:500 dilution.

Techniques: Incubation, SDS Page, Western Blot, Control

Journal: Molecular and Cellular Biology

Article Title: Human Insulin Growth Factor 2 mRNA Binding Protein 2 Increases MicroRNA 33a/b Inhibition of Liver ABCA1 Expression and Alters Low-Density Apolipoprotein Levels in Mice

doi: 10.1128/MCB.00058-20

Figure Lengend Snippet: Both AGO2 and STAU1 are required for IGF2BP2-dependent miR-33a/b ABCA1 silencing. (A) qRT-PCR analysis was used to determine relative endogenous ABCA1 mRNA expression in miR-33a/b-expressing cells treated with siRNAs targeting the indicated genes (indicated by “si” suffix). ABCA1 mRNA expression values were compared to GAPDH mRNA expression, which was set at 1. (B) ABCA1 protein level was determined in miR-33a/b-expressing cells treated with the indicated siRNA. Actin was used as a loading control. (C) qRT-PCR analysis was used to determine the relative endogenous ABCA1 mRNA expression in miR-33a/b-expressing cells transfected with pCMV plasmids expressing the indicated genes (OE). (D) ABCA1 protein level was determined in miR-33a/b-expressing cells transfected with pCMV plasmids expressing the indicated genes (OE). Con, control. **, P < 0.001; ***, P < 0.0001. The qRT-PCR data are the averages from five independent experiments. Values are the means ± SEM. The Western blot panels represent the average results of five independent experiments.

Article Snippet: Anti-ABCA1 polyclonal antibodies were from Abcam (ab1880) and used at a 1:500 dilution; anti-IGF2BP1 monoclonal antibodies were from Santa Cruz (IGF2BP1 [sc-390149], IGF2BP2 [sc-377014], and IGF2BP3 [sc-365640]), and all were used at a 1:2,000 dilution; anti-AGO2 polyclonal antibodies were from Abcam (ab32381) and used at a 1:750 dilution; anti-STAU1 polyclonal antibodies were from Sigma-Aldrich (SAB1406490) and used at a 1:1,000 dilution; anti-G3BP polyclonal antibodies were from Thermo Fisher (G3BP1 [PA5-29455]; G3BP2 [PA5-53797]) and were used at a 1:500 dilution.

Techniques: Quantitative RT-PCR, Expressing, Control, Transfection, Western Blot

Journal: Molecular and Cellular Biology

Article Title: Human Insulin Growth Factor 2 mRNA Binding Protein 2 Increases MicroRNA 33a/b Inhibition of Liver ABCA1 Expression and Alters Low-Density Apolipoprotein Levels in Mice

doi: 10.1128/MCB.00058-20

Figure Lengend Snippet: Overexpression of IGF2BP2 cannot restore miR-33a/b-dependent ABCA1 silencing in cells lacking AGO2 or STAUI1 expression. (A) qRT-PCR analysis was used to determine relative endogenous ABCA1 mRNA expression in miR-33a/b-expressing cells transfected with pCMV-IGF2BP2 (OE) and treated with siRNAs targeting the indicated genes (“si” suffix). ABCA1 mRNA expression values were compared to GAPDH mRNA expression. (B) ABCA1 protein level was determined in miR-33a/b-expressing cells transfected with pCMV-IGF2BP2 (OE) and treated with the indicated siRNAs. Actin was used as a loading control. (C) qRT-PCR analysis was used to determine relative endogenous ABCA1 mRNA expression in miR-33a/b-expressing cells transfected with pCMV plasmids expressing the indicated genes (OE) and treated with IGF2BP2 siRNA. (D) ABCA1 protein level was determined in miR-33a/b-expressing cells transfected with pCMV plasmids expressing the indicated genes (OE) and treated with IGF2BP2 siRNA. Actin was used as a loading control. Con, control. **, P < 0.001; ***, P < 0.0001. The qRT-PCR data represent the average results of five independent experiments. Values are the means ± SEM. The Western blot panels represent the average results of five independent experiments.

Article Snippet: Anti-ABCA1 polyclonal antibodies were from Abcam (ab1880) and used at a 1:500 dilution; anti-IGF2BP1 monoclonal antibodies were from Santa Cruz (IGF2BP1 [sc-390149], IGF2BP2 [sc-377014], and IGF2BP3 [sc-365640]), and all were used at a 1:2,000 dilution; anti-AGO2 polyclonal antibodies were from Abcam (ab32381) and used at a 1:750 dilution; anti-STAU1 polyclonal antibodies were from Sigma-Aldrich (SAB1406490) and used at a 1:1,000 dilution; anti-G3BP polyclonal antibodies were from Thermo Fisher (G3BP1 [PA5-29455]; G3BP2 [PA5-53797]) and were used at a 1:500 dilution.

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Transfection, Control, Western Blot

Journal: Molecular and Cellular Biology

Article Title: Human Insulin Growth Factor 2 mRNA Binding Protein 2 Increases MicroRNA 33a/b Inhibition of Liver ABCA1 Expression and Alters Low-Density Apolipoprotein Levels in Mice

doi: 10.1128/MCB.00058-20

Figure Lengend Snippet: Overexpression of hIGF2BP2 in mice has a slight effect on fatty acid composition and C16/0/C18:0 ratio. (A) Total liver fatty acid compositions from the three cohorts were determined by analyzing fatty acid methyl esters by GC. Black bars, CMV-GFP vector control cohort; white bars, CMV-IGF2BP2 cohort; gray bars, CMV-miR-33a/b cohort. (B) The C16/C18 ratio was determined by using the percentage of total C16:0 and C18:0 fatty acids.

Article Snippet: Anti-ABCA1 polyclonal antibodies were from Abcam (ab1880) and used at a 1:500 dilution; anti-IGF2BP1 monoclonal antibodies were from Santa Cruz (IGF2BP1 [sc-390149], IGF2BP2 [sc-377014], and IGF2BP3 [sc-365640]), and all were used at a 1:2,000 dilution; anti-AGO2 polyclonal antibodies were from Abcam (ab32381) and used at a 1:750 dilution; anti-STAU1 polyclonal antibodies were from Sigma-Aldrich (SAB1406490) and used at a 1:1,000 dilution; anti-G3BP polyclonal antibodies were from Thermo Fisher (G3BP1 [PA5-29455]; G3BP2 [PA5-53797]) and were used at a 1:500 dilution.

Techniques: Over Expression, Plasmid Preparation, Control

Journal: Molecular and Cellular Biology

Article Title: Human Insulin Growth Factor 2 mRNA Binding Protein 2 Increases MicroRNA 33a/b Inhibition of Liver ABCA1 Expression and Alters Low-Density Apolipoprotein Levels in Mice

doi: 10.1128/MCB.00058-20

Figure Lengend Snippet: Overexpressing IGF2BP2 in mice results in loss of ABCA1 expression, elevation in serum cholesterol, and defects in apolipoprotein homeostasis. (A) GFP-IGF2BP2 liver protein levels in mice injected with control CMV-GFP (Ad-CMV-GFP) or CMV-GFP-IGF2PB2 (Ad-CMV-GFP-IGF2BP2) were determined. Actin was used as a loading control. (B) qRT-PCR analysis was used to determine the relative endogenous ABCA1 mRNA expression levels in mice injected with control CMV-GFP (Ad-CMV-GFP) or CMV-GFP-IGF2PB2 (Ad-CMV-GFP-IGF2BP2). (C) Serum cholesterol, triglyceride, and apolipoprotein levels were determined as described in the legend to Fig. 7. Filled circles, Ad-CMV-GFP; open circles, Ad-CMV-GFP-IGF2BP2. (D) Apolipoprotein levels were determined using the Lipoprint assay. Filled circles, Ad-CMV-GFP; open circles, Ad-CMV-GFP-IGF2BP2. **, P < 0.001; ***, P < 0.0001. (E) Liver enzymes activities were determined. ALP, alkaline phosphatase; AST, aspartate aminotransferase; ALT, alanine aminotransferase. Values are the means ± SEM (n = 3).

Article Snippet: Anti-ABCA1 polyclonal antibodies were from Abcam (ab1880) and used at a 1:500 dilution; anti-IGF2BP1 monoclonal antibodies were from Santa Cruz (IGF2BP1 [sc-390149], IGF2BP2 [sc-377014], and IGF2BP3 [sc-365640]), and all were used at a 1:2,000 dilution; anti-AGO2 polyclonal antibodies were from Abcam (ab32381) and used at a 1:750 dilution; anti-STAU1 polyclonal antibodies were from Sigma-Aldrich (SAB1406490) and used at a 1:1,000 dilution; anti-G3BP polyclonal antibodies were from Thermo Fisher (G3BP1 [PA5-29455]; G3BP2 [PA5-53797]) and were used at a 1:500 dilution.

Techniques: Expressing, Injection, Control, Quantitative RT-PCR

Journal: Molecular and Cellular Biology

Article Title: Human Insulin Growth Factor 2 mRNA Binding Protein 2 Increases MicroRNA 33a/b Inhibition of Liver ABCA1 Expression and Alters Low-Density Apolipoprotein Levels in Mice

doi: 10.1128/MCB.00058-20

Figure Lengend Snippet: Mice expressing GFP-IGF2BP2 accumulate mature forms of SREBP1/2 and have elevated lipid protein expression. Protein levels were determined by Western analysis in mice injected with CMV-GFP (Ad-CMV-GFP) or CMV-GFP-IGF2BP2 (Ad-CMV-GFP-IGF2BP2). Actin was used as a loading control.

Article Snippet: Anti-ABCA1 polyclonal antibodies were from Abcam (ab1880) and used at a 1:500 dilution; anti-IGF2BP1 monoclonal antibodies were from Santa Cruz (IGF2BP1 [sc-390149], IGF2BP2 [sc-377014], and IGF2BP3 [sc-365640]), and all were used at a 1:2,000 dilution; anti-AGO2 polyclonal antibodies were from Abcam (ab32381) and used at a 1:750 dilution; anti-STAU1 polyclonal antibodies were from Sigma-Aldrich (SAB1406490) and used at a 1:1,000 dilution; anti-G3BP polyclonal antibodies were from Thermo Fisher (G3BP1 [PA5-29455]; G3BP2 [PA5-53797]) and were used at a 1:500 dilution.

Techniques: Expressing, Western Blot, Injection, Control

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 physically associates with IGF2BP2 in macrophages. (A) LC–MS/MS summary of proteins enriched by circSMAD4 RNA pull-down. (B) Western blot validation of IGF2BP2 in circSMAD4 sense (vs antisense) pull-down from TC-hMDMs. (C) IGF2BP2 RIP–qPCR showing circSMAD4 enrichment over IgG in TC-hMDMs. (D–E) catRAPID prediction and ViennaRNA RNAfold secondary-structure modeling indicating multiple candidate IGF2BP2-binding regions on circSMAD4. (F) Western blot of IGF2BP2 after pull-down with circSMAD4 fragments (1#–3#). (G) Schematic of IGF2BP2 domain architecture and the Flag-tagged truncation/deletion constructs used for mapping circSMAD4 interaction (designed based on catRAPID prediction and annotated RRM/KH domain boundaries). (H) Anti-Flag RIP–qPCR showing circSMAD4 enrichment precipitated by the indicated Flag-tagged IGF2BP2 truncation/deletion constructs (presented as % input and normalized to IgG). (I) Nuclear–cytoplasmic fractionation followed by RT–qPCR showing circSMAD4 distribution and the effect of IGF2BP2 knockdown on the nuclear-to-cytoplasmic ratio of circSMAD4 in TC-hMDMs. Fractionation quality was validated using nuclear/cytoplasmic marker transcripts/proteins. (J) Representative immunofluorescence/ISH images showing circSMAD4 signals and IGF2BP2 staining in macrophages (CD163) with nuclear counterstaining (DAPI). Scale bar, 50 μm. (K–N) qPCR and Western blot showing no reciprocal change in expression between circSMAD4 and IGF2BP2 upon knockdown/overexpression. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Gene silencing was performed using lentiviral shRNAs. shRNAs targeting the human circSMAD4 back-splice junction (and the murine circSmad4 ortholog, avoiding linear Smad4) as well as IGF2BP2 were cloned into the pLKO.1-puro vector (Addgene, #8453).

Techniques: Liquid Chromatography with Mass Spectroscopy, Western Blot, Biomarker Discovery, Binding Assay, Construct, Fractionation, Quantitative RT-PCR, Knockdown, Marker, Immunofluorescence, Staining, Expressing, Over Expression

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 facilitates IGF2BP2-dependent stabilization of m6A-marked transcripts. (A) Venn diagram intersecting ENCORI-predicted IGF2BP2 targets with DEGs from shIGF2BP2 versus shNC and shcircSMAD4 versus shNC mRNA-seq, identifying shared candidates. (B) MeRIP–qPCR showing m6A enrichment on COL4A1, SPI1, and ACTA2 candidate regions (CRDs) in shNC and shIGF2BP2 cells. (C) IGF2BP2-RIP–qPCR showing IGF2BP2 binding to COL4A1, SPI1, and ACTA2 CRDs in shNC + Vector, shcircSMAD4 + Vector, shNC + IGF2BP2, and shcircSMAD4 + IGF2BP2 groups. (D) Biotin-circSMAD4 pull-down followed by qPCR showing enrichment of COL4A1, SPI1, and ACTA2 CRDs in Vector + shNC, circSMAD4 + shNC, Vector + shIGF2BP2, and circSMAD4 + shIGF2BP2 groups. (E–G) Schematics of m6A-site mutations introduced into COL4A1, SPI1, and ACTA2 reporters. (H–J) Dual-luciferase assays for CRD reporters (WT and m6A-mutant) in Vector, circSMAD4, and IGF2BP2 groups. (K–M) MeRIP–qPCR for WT and m6A-mutant CRD reporters in Vector, circSMAD4, and IGF2BP2 groups. (N–P) mRNA decay assays of endogenous COL4A1, SPI1, and ACTA2 following circSMAD4 knockdown with Vector or IGF2BP2 overexpression. Half-life estimated by one-phase decay (Y0 = 1, Plateau = 0). (Q–S) mRNA decay assays of endogenous COL4A1, SPI1, and ACTA2 following circSMAD4 overexpression with shNC or shIGF2BP2. Half-life estimated by one-phase decay (Y0 = 1, Plateau = 0). ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Gene silencing was performed using lentiviral shRNAs. shRNAs targeting the human circSMAD4 back-splice junction (and the murine circSmad4 ortholog, avoiding linear Smad4) as well as IGF2BP2 were cloned into the pLKO.1-puro vector (Addgene, #8453).

Techniques: Binding Assay, Plasmid Preparation, Luciferase, Mutagenesis, Knockdown, Over Expression

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: Proposed model: circSMAD4 drives matrix-remodeling TAM programs in LUAD. Schematic summary illustrating that circSMAD4 in tumor-associated macrophages promotes a matrix-remodeling, M2-like state through two post-transcriptional routes: (i) circSMAD4 sequesters miR-562 in an AGO2-dependent manner to relieve repression of COL4A1 mRNA; (ii) circSMAD4 associates with IGF2BP2 to enhance the stability of m6A-marked transcripts, including COL4A1, SPI1, and ACTA2 (α-SMA). These combined outputs reinforce extracellular matrix remodeling within the LUAD tumor microenvironment.

Article Snippet: Gene silencing was performed using lentiviral shRNAs. shRNAs targeting the human circSMAD4 back-splice junction (and the murine circSmad4 ortholog, avoiding linear Smad4) as well as IGF2BP2 were cloned into the pLKO.1-puro vector (Addgene, #8453).

Techniques:

Journal: Discover Oncology

Article Title: Novel N 6 -methyladenosine (m 6 A) writer METTL16 promotes the cervical cancer tumorigenesis by targeting FTH1-dependent ferroptosis

doi: 10.1007/s12672-026-04403-8

Figure Lengend Snippet: IGF2BP2 promoted the stability of FTH1 mRNA. A The immunofluorescent staining showed the subcellular localization of IGF2BP2 and FTH1 in HeLa cells. B The positive correlation within IGF2BP2 and FTH1 in clinical samples ( http://gepia.cancer-pku.cn/index.html ). C RNA immunoprecipitation (RIP) assay showed the immunoprecipitated FTH1 mRNA by anti-IGF2BP2 antibody. D RNA decay analysis reported the stability of FTH1 mRNA in cervical cancer cells with IGF2BP2 overexpression. E RNA decay analysis reported the stability of FTH1 mRNA in cervical cancer cells with METTL16 silencing (sh-METTL16-1, sh-METTL16-2). F RNA decay analysis reported the stability of FTH1 mRNA in cervical cancer cells with IGF2BP2 silencing (si-IGF2BP2) and METTL16 overexpression. * p < 0.05, ** p < 0.01

Article Snippet: Anti-METTL16 antibody (1:2000, 19924-1-AP, Proteintech), anti-FTH1 antibody (1:1000, 83428-1-RR, Proteintech), anti-IGF2BP2 antibody (1:1000, 11601-1-AP, Proteintech) acted as the primary antibody for incubation.

Techniques: Staining, RNA Immunoprecipitation, Immunoprecipitation, Over Expression

Journal: The Egyptian Journal of Biochemistry and Molecular Biology

Article Title: Insulin-like Growth Factor 2 Binding Protein 2 Gene Polymorphism in Egyptian Patients with Type 2 Diabetes

doi: 10.21608/ejb.2018.19873

Figure Lengend Snippet: Figure 1. Agarose gel electrophoresis of restriction digests of PCR products of IGF2BP2 gene using MboII restriction enzyme on 2%(w/v) agarose gel. M:50 bp Marker, lanes 3, 5 and 8 are GG genotypes, lanes 1, 6 and 7are GT genotypes and lanes 2 and 4 are TT genotypes for T2DM patients. Lanes11, 14 and 15 are GG genotypes, lanes 9, 10, 12 and 13 are GT genotypes for controls.

Article Snippet: Fasting serum insulin (DRG Insulin ELISA kit, DRG International, Inc., USA) and IGF2BP2 protein (IGF2BP2 ELISA Kit, Cusabio Biotech Co., Ltd, China) levels were determined according to manufacturer’s protocol and measured by using microplate reader (Stat Fax 2100, Awareness Technology, Inc., USA).

Techniques: Agarose Gel Electrophoresis, Marker

Journal: Fertility and sterility

Article Title: HMGA2-mediated tumorigenesis through angiogenesis in leiomyoma

doi: 10.1016/j.fertnstert.2020.05.036

Figure Lengend Snippet: HMGA2 promotes angiogenesis and mediates IGF2BP2/pAKT activity in human umbilical vein endothelial cell (HUVEC) migration. (A) Tube formation of HUVEC in the CM from LM and MM cells without (control) and with HMGA2 overexpression. (B) Histogram demonstrating the number of complete tubes and significance analysis. (C) Histogram showing the number of migrated HUVEC (Transwell assay) in the CM from LM and MM cells without (control) and with HMGA2 overexpression. (D) Histogram showing the relative migration rate of HUVEC after scratching in the CM from LM and MM cells without (control) and with HMGA2 overexpression. *P<.05; **P<.01. (E) Western blot analysis illustrating pAKT and IGF2BP2 expression in LM and MM cells with and without HMGA2 overexpression. (F) Expression analysis of HMGA2 and pAKT in LM and MM cells after silencing IGF2BP2 expression. (G, H) HUVEC migration analysis in the CM from LM and MM cells without (control) and with HMGA2 overexpression, as well as silencing IGF2BP2.

Article Snippet: IGF2BP2 siRNA and control siRNA were purchased from Santa Cruz and used for transfecting cells with Lipofectamine 3000 (Invitrogen) according to the manufacturer’s protocol.

Techniques: Activity Assay, Migration, Control, Over Expression, Transwell Assay, Western Blot, Expressing

Journal: Cell Death and Differentiation

Article Title: IGF2BP2 regulates DANCR by serving as an N6-methyladenosine reader

doi: 10.1038/s41418-019-0461-z

Figure Lengend Snippet: a DANCR promotes the tumor growth of BXPC-3 cells. DANCR overexpression (DANCR) and vector control (pCDH-MSCV-copGFP-T2A-Pu) cells were subcutaneously injected into the nude mice with 1 × 10 4 , 1 × 10 5 , 5 × 10 5 cells per mouse as indicated in “Materials and methods”. b DANCR KO significantly suppresses the tumor growth rate of BXPC-3 cells. DANCR KO and vector control (pY108) cells were subcutaneously injected into the nude mice with 1 × 10 4 , 1 × 10 5 , 5 × 10 5 cells per mouse as in ( a ). ( c ) DANCR KO inhibits tumorigenesis of BXPC-3 cells in orthotopic pancreatic tumor mouse model. DANCR KO and vector control cells were injected into the pancreas of nude mice with 1 × 10 6 cells per mouse as detailed in “Materials and methods”. Values in a , b are mean ± SEM. P < 0.05; ** P < 0.01.

Article Snippet: The following constructs were obtained from Addgene (Watertown, MA): pY108 (lenti-AsCpf1, #84739); LentiCRISPR v2 (#52961); MSCV-human Igf2bp2-IRES-GFG (#91890). pCDH-MSCV-copGFP-T2A-Pu was purchased from System Biosciences (Mountain View, CA).

Techniques: Over Expression, Plasmid Preparation, Injection