primary antibody against igf1r  (Thermo Fisher)

Journal: Frontiers in Genetics

Article Title: Circ-MFN2 Positively Regulates the Proliferation, Metastasis, and Radioresistance of Colorectal Cancer by Regulating the miR-574-3p/IGF1R Signaling Axis

doi: 10.3389/fgene.2021.671337

Figure Lengend Snippet: The primer sequences used for qRT-PCR.

Article Snippet: After blocked with 5% non-fat milk, the membranes were hatched with primary antibodies against IGF1R (1:2,000, Bioss, Beijing, China), MFN2 (1:1,000, Bioss) or GAPDH (1:10,000, Bioss).

Techniques:

Journal: Frontiers in Genetics

Article Title: Circ-MFN2 Positively Regulates the Proliferation, Metastasis, and Radioresistance of Colorectal Cancer by Regulating the miR-574-3p/IGF1R Signaling Axis

doi: 10.3389/fgene.2021.671337

Figure Lengend Snippet: MiR-574-3p could target IGF1R. (A) The sequences of IGF1R 3′UTR-WT and IGF1R 3′UTR-MUT were shown. (B,C) The interaction between IGF1R 3′UTR and miR-574-3p was confirmed using dual-luciferase reporter assay. (D,E) The mRNA and protein levels of IGF1R in CRC tumor tissues and non-cancer tissues were measured using qRT-PCR and WB analysis. (F,G) QRT-PCR and WB analysis were used to detect the mRNA and protein levels of IGF1R in CRC cell lines (SW620 and SW480) and NCM460 cells. (H) The correlation between IGF1R and miR-574-3p was analyzed using Pearson correlation analysis. * P < 0.05.

Article Snippet: After blocked with 5% non-fat milk, the membranes were hatched with primary antibodies against IGF1R (1:2,000, Bioss, Beijing, China), MFN2 (1:1,000, Bioss) or GAPDH (1:10,000, Bioss).

Techniques: Luciferase, Reporter Assay, Quantitative RT-PCR

Journal: Frontiers in Genetics

Article Title: Circ-MFN2 Positively Regulates the Proliferation, Metastasis, and Radioresistance of Colorectal Cancer by Regulating the miR-574-3p/IGF1R Signaling Axis

doi: 10.3389/fgene.2021.671337

Figure Lengend Snippet: The regulation of miR-574-3p mimic and IGF1R overexpression on CRC progression. SW620 and SW480 cells were transfected with miR-NC, miR-574-3p, miR-574-3p + pcDNA or miR-574-3p + IGF1R, and non-transfected cells were used as control. (A) The protein level of IGF1R was measured by WB analysis to evaluate transfection efficiency. (B,C) The viability of cells was assessed using MTT assay. (D) Colony formation assay was determined to evaluate the number of colonies. (E,F) The migration and invasion of cells were detected using transwell assay. (G,H) The survival fraction of cells was examined using colony formation assay. * P < 0.05.

Article Snippet: After blocked with 5% non-fat milk, the membranes were hatched with primary antibodies against IGF1R (1:2,000, Bioss, Beijing, China), MFN2 (1:1,000, Bioss) or GAPDH (1:10,000, Bioss).

Techniques: Over Expression, Transfection, MTT Assay, Colony Assay, Migration, Transwell Assay

Journal: Frontiers in Genetics

Article Title: Circ-MFN2 Positively Regulates the Proliferation, Metastasis, and Radioresistance of Colorectal Cancer by Regulating the miR-574-3p/IGF1R Signaling Axis

doi: 10.3389/fgene.2021.671337

Figure Lengend Snippet: Circ-MFN2 and miR-574-3p regulated IGF1R expression. SW620 and SW480 cells were transfected with si-NC, si-circ-MFN2, si-circ-MFN2 + anti-miR-NC or si-circ-MFN2 + anti-miR-574-3p, and non-transfected cells were used as control. The protein level of IGF1R in SW620 (A) and SW480 (B) cells was measured using WB analysis. * P < 0.05.

Article Snippet: After blocked with 5% non-fat milk, the membranes were hatched with primary antibodies against IGF1R (1:2,000, Bioss, Beijing, China), MFN2 (1:1,000, Bioss) or GAPDH (1:10,000, Bioss).

Techniques: Expressing, Transfection

Journal: The FASEB Journal

Article Title: Impaired myoblast differentiation and muscle IGF‐1 receptor signaling pathway activation after N‐glycosylation inhibition

doi: 10.1096/fj.202400213rr

Figure Lengend Snippet: FIGURE 4 IGF-1 production and IGF1R signaling pathway activation in TA muscle of WT and MLC/mIgf-1 mice treatment with low TUN dose (0.1 mg/kg of TUN for 15 days). (A) Expression of endogenous Igf-1 mRNA isoforms (i.e., Igf-1Ea and Igf-1Ec mRNAs) and total Igf-1 (i.e., endogenous and mIgf-1 transgene mRNAs) in the TA muscle of WT (n = 3) and MLC/mIgf-1 (n = 3) mice treated with TUN. (B) Representative immunoblotting and (C) band densitometry analysis of TA muscle of WT and MLC/mIgf1 mice using an antibody directed against mature mouse IGF-1 sequence. Three bands at a molecular weight of ~22, ~17, and ~12 kDa were detected in TA muscle of MLC/ mIgf1 mice. Recombinant mouse IGF-1 mature protein was loaded as a positive control for mature IGF-1 (~7 kDa). (D) Representative immunoblotting and (E) band densitometry analysis of TA muscle of WT and MLC/mIgf1 mice using antibodies directed against phosphorylated (pIGF1R) and total IGF1R, phosphorylated (pAKT) and total AKT, and phosphorylated (pERK1/2) and total ERK1/2. All bar charts are presented as mean values ± SD. Significant differences were determined using unpaired t-test (C) or two-way ANOVA followed by Tukey's multiple comparison post hoc tests (A, E). *Significantly different compared to CTR; #Significantly different compared to WT mice; * and # p ≤ .05; ** p ≤ .01 *** and ### p ≤ .001.

Article Snippet: Primary antibodies against phospho- IGF1R β (1:2000; cat. n. 3024 Cell Signaling Technology), total IGF1R β (1:2000; cat. n. 3027 Cell Signaling Technology), phospho- Akt (Ser473) (1:2000; cat. n. 9271 Cell Signaling Technology), total Akt (1:2000; cat. n. 9272 Cell Signaling Technology), phospho- p44/42 (ERK1/2) (1:2000; cat. n. 9101 Cell Signaling Technology), total p44/42 (ERK1/2) (1:2000; cat. n. 9102 Cell Signaling Technology), PCNA (1:5000, cat. n. MAB 424R Millipore), MF20 (1:500, DSHB), and mouse/rat biotinylated IGF- 1 antibody (1:300, cat. n. BAF791 R&D Systems) were incubated overnight at 4°C.

Techniques: Activation Assay, Expressing, Western Blot, Sequencing, Molecular Weight, Recombinant, Positive Control, Comparison

Journal: The FASEB Journal

Article Title: Impaired myoblast differentiation and muscle IGF‐1 receptor signaling pathway activation after N‐glycosylation inhibition

doi: 10.1096/fj.202400213rr

Figure Lengend Snippet: FIGURE 5 Effect of TUN treatment on IGF1R production and IGF1R signaling pathway activation in C2C12. (A) IGF1R and IGF1R proreceptor production in C2C12 cells treated with 0.01 μg/mL of TUN and harvested at day 1. (B) Immunofluorescence analysis of C2C12 cells stained with ant-IGF1R and DAPI, the scale bar represents 200 μm. (C) Representative immunoblotting and (D) band densitometry analysis of IGF-1-induced IGF-1R signaling pathway activation in CTR- and TUN-treated C2C12 cells using antibodies directed against phosphorylated (pIGF1R) and total IGF1R, phosphorylated (pAKT) and total AKT, and phosphorylated (pERK1/2) and total ERK1/2. The calculation of pIGF1R level was normalized against total lysed protein instead of total IGF1R to account for the marked reduction of total IGF1R after TUN treatment. All bar charts are presented as mean values ± SD. Significant differences were determined using unpaired t-test (A) or one-way ANOVA followed by Tukey's multiple comparison post hoc tests (D). *Significantly different compared to CTR, #Significantly different compared to IGF-1-treated cells; ** p ≤ .01, *** p ≤ .001; ## p ≤ .01, ### p ≤ .001.

Article Snippet: Primary antibodies against phospho- IGF1R β (1:2000; cat. n. 3024 Cell Signaling Technology), total IGF1R β (1:2000; cat. n. 3027 Cell Signaling Technology), phospho- Akt (Ser473) (1:2000; cat. n. 9271 Cell Signaling Technology), total Akt (1:2000; cat. n. 9272 Cell Signaling Technology), phospho- p44/42 (ERK1/2) (1:2000; cat. n. 9101 Cell Signaling Technology), total p44/42 (ERK1/2) (1:2000; cat. n. 9102 Cell Signaling Technology), PCNA (1:5000, cat. n. MAB 424R Millipore), MF20 (1:500, DSHB), and mouse/rat biotinylated IGF- 1 antibody (1:300, cat. n. BAF791 R&D Systems) were incubated overnight at 4°C.

Techniques: Activation Assay, Immunofluorescence, Staining, Western Blot, Comparison

Journal: Oncology Letters

Article Title: Receptor tyrosine kinase expression in high-grade gliomas before and after chemoradiotherapy

doi: 10.3892/ol.2019.11017

Figure Lengend Snippet: Significantly changed receptor tyrosine kinases between initial and recurrent gliomas.

Article Snippet: 5% skim milk was used to block the membrane at room temperature for 1 h. Primary rabbit polyclonal antibodies against IGF1R (ProteinTech Group, Inc.; cat. no. 10297-1-AP; dilution, 1:200) were used at 4°C overnight.

Techniques:

Journal: Oncology Letters

Article Title: Receptor tyrosine kinase expression in high-grade gliomas before and after chemoradiotherapy

doi: 10.3892/ol.2019.11017

Figure Lengend Snippet: IGFR was activated by TMZ. (A) Western blot analysis revealed that IGF1R expression was increased in glioma cells after treatment with TMZ. Expression levels was normalized by the level of GAPDH and expressed as fold-changes of those in parental cells. The data are presented as mean ± SD. ***P<0.001 as indicated. (B) Data analysis of 135 initial and 47 recurrent high-grade glioma cases revealed that IGFR signaling was highly activated in recurrent gliomas. TMZ, temozolomide; IGF1R, insulin-like growth factor 1 receptor.

Article Snippet: 5% skim milk was used to block the membrane at room temperature for 1 h. Primary rabbit polyclonal antibodies against IGF1R (ProteinTech Group, Inc.; cat. no. 10297-1-AP; dilution, 1:200) were used at 4°C overnight.

Techniques: Western Blot, Expressing