Journal: Experimental Hematology & Oncology

Article Title: Physical activity improves outcomes of combined lenvatinib plus anti-PD-1 therapy in unresectable hepatocellular carcinoma: a retrospective study and mouse model

doi: 10.1186/s40164-022-00275-0

Figure Lengend Snippet: Immunohistochemical staining and analysis of subcutaneous tumors. Immunohistochemical staining was conducted on subcutaneous tumor tissue microarray. H-score were calculated to quantify the expression levels of marker proteins. Representative images captured at 40 ×. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Foxp3 forkhead box protein p3, CTLA4 cytotoxic T-lymphocyte-associated protein 4, TIGIT T cell immunoreceptor with Ig and ITIM domains, TIM3 T-cell immunoglobulin mucin-3, VISTA V domain-containing Ig suppressor of T-cell activation

Article Snippet: Primary Abs used for immunohistochemical staining included: anti-CD4 Ab (#25229, CST), anti-CD8α Ab (#98941,CST), anti-Forkhead box protein p3 (Foxp3) Ab (ab253297, Abcam), anti-F4/80 Ab (#70076, CST), anti-CTLA4 Ab (A00020, Boster), anti-T cell immunoreceptor with Ig and immunoreceptor tyrosine-based inhibitory motif (ITIM) domains (TIGIT) Ab (A01962, Boster), anti-TIM3 Ab (ab241332, Abcam) and, anti-V domain-containing Ig suppressor of T-cell activation (VISTA) Ab (#54979, CST).

Techniques: Immunohistochemical staining, Staining, Microarray, Expressing, Marker, Activation Assay

Journal: Journal of Functional Foods

Article Title: Noni (Morinda citrifolia L.) fruit polysaccharide ameliorated high-fat diet-induced obesity by modulating gut microbiota and improving bile acid metabolism

doi: 10.1016/j.jff.2023.105408

Figure Lengend Snippet: Fig. 2. Effect of NFP on bile acids synthesis in mice under a high-fat diet. (A) The content of TBA in the liver. (B) The expression of CYP7A1 in the liver. (C) Relative protein expression levels of liver FXR. (D) Relative protein expression levels of liver SHP. (E) Bargraph of liver bile acid-related protein expression. (F) Bargraph of intestinal bile acid-related protein expression. (G) Relative protein expression levels of colonic FXR. (H) Relative expression levels of colonic FGF15. (I) Relative expression levels of colonic TGR5. Data are shown as mean ± SEM (n = 5); *p < 0.05, **p < 0.01 versus the NC group; #p < 0.05, ##p < 0.01 versus the HFD group.

Article Snippet: The antibodies GAPDH (BA2913), SHP (A03866-1), and TGR5 (A01958-1) for Western blot detection were purchased from Boster Bioengineering Co., Ltd. (California, USA).

Techniques: Expressing

Journal: BMC Veterinary Research

Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)

doi: 10.1186/s12917-018-1695-6

Figure Lengend Snippet: Increasing chi-miR-3031 and siRNA-IGFBP5 levels notably decreased IGFBP5 mRNA expression. a : Effect of chi-miR-3031 on IGFBP5 mRNA. b : Effect of siRNA-IGFBP5 on IGFBP5 mRNA. P < 0.01 is shown as **. MC: Chi-miR-3031 mimic. NC: Negative control

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with a rabbit anti-IGFBP5 polyclonal antibody (1:300, Boster, USA), a rabbit anti-mTOR polyclonal antibody (1:100, Boster, USA), a rabbit anti-p-mTOR polyclonal antibody (1:100, Boster, USA) or a mouse β-actin monoclonal primary antibody (1:1000, Beyotime, Jiangsu, China).

Techniques: Expressing, Negative Control

Journal: BMC Veterinary Research

Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)

doi: 10.1186/s12917-018-1695-6

Figure Lengend Snippet: Effects of chi-miR-3031 and siRNA-IGFBP5 on IGFBP5 protein levels. a : Western blot analysis results. b : Densitometric quantification of western blot results. Protein levels were normalized to β-actin. P < 0.05 is shown as *, and P < 0.01 is shown as **. MC: Chi-miR-3031 mimic. NC: Negative control

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with a rabbit anti-IGFBP5 polyclonal antibody (1:300, Boster, USA), a rabbit anti-mTOR polyclonal antibody (1:100, Boster, USA), a rabbit anti-p-mTOR polyclonal antibody (1:100, Boster, USA) or a mouse β-actin monoclonal primary antibody (1:1000, Beyotime, Jiangsu, China).

Techniques: Western Blot, Negative Control

Journal: BMC Veterinary Research

Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)

doi: 10.1186/s12917-018-1695-6

Figure Lengend Snippet: Protein expression levels of κ-casein ( a ) and β-casein ( b ) in GMECs transfected with MC, NC and siRNA-IGFBP5. P < 0.05 is shown as *, and P < 0.01 is shown as **. MC: Chi-miR-3031 mimic. NC: Negative control

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with a rabbit anti-IGFBP5 polyclonal antibody (1:300, Boster, USA), a rabbit anti-mTOR polyclonal antibody (1:100, Boster, USA), a rabbit anti-p-mTOR polyclonal antibody (1:100, Boster, USA) or a mouse β-actin monoclonal primary antibody (1:1000, Beyotime, Jiangsu, China).

Techniques: Expressing, Transfection, Negative Control

Journal: BMC Veterinary Research

Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)

doi: 10.1186/s12917-018-1695-6

Figure Lengend Snippet: Effects of chi-miR-3031 and siRNA-IGFBP5 on p-mTOR protein levels. a : Western blot analysis results. b : Densitometric quantification of western blot results. P < 0.05 is shown as *, and P < 0.01 is shown as **. MC: Chi-miR-3031 mimic. NC: Negative control

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with a rabbit anti-IGFBP5 polyclonal antibody (1:300, Boster, USA), a rabbit anti-mTOR polyclonal antibody (1:100, Boster, USA), a rabbit anti-p-mTOR polyclonal antibody (1:100, Boster, USA) or a mouse β-actin monoclonal primary antibody (1:1000, Beyotime, Jiangsu, China).

Techniques: Western Blot, Negative Control

Journal: BMC Veterinary Research

Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)

doi: 10.1186/s12917-018-1695-6

Figure Lengend Snippet: Primer information for RT-qPCR

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with a rabbit anti-IGFBP5 polyclonal antibody (1:300, Boster, USA), a rabbit anti-mTOR polyclonal antibody (1:100, Boster, USA), a rabbit anti-p-mTOR polyclonal antibody (1:100, Boster, USA) or a mouse β-actin monoclonal primary antibody (1:1000, Beyotime, Jiangsu, China).

Techniques: Sequencing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: The effects of HIF-1alpha on gene expression profiles of NCI-H446 human small cell lung cancer cells

doi: 10.1186/1756-9966-28-150

Figure Lengend Snippet: 65 genes upregulated by HIF-1alpha more than 2.0-fold in three pairwise comparisons

Article Snippet: The rabbit anti-human primary antibodies (Wuhan Boster Biological Engineering Technology Limited Company) that detect IGFBP5, SOCS1, IL-6 and STAT3(Signal Transducer and Activator of Transcription 3)were incubated with membranes overnight at 4°C.

Techniques: Transduction, Binding Assay, Derivative Assay, De-Phosphorylation Assay, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: The effects of HIF-1alpha on gene expression profiles of NCI-H446 human small cell lung cancer cells

doi: 10.1186/1756-9966-28-150

Figure Lengend Snippet: Real-time PCR analysis of upregulated or downregulated gene expression in response to HIF-1alpha (A) Aliquots of the same RNA preparations used for microarray hybridization were analyzed by quantitative real-time PCR . In three pairwise comparisons, the upregulation-folds of IGFBP5, IRS4, TNFAIP6, SOCS1, IL-6, VEGF-A mRNA expression were calculated. The mean and standard error are shown (p < 0.05). (B) Aliquots of the same RNA preparations used for microarray hybridization were analyzed by quantitative real-time PCR. In three pairwise comparisons, the downregulation-folds of IGFBP3, ZNF569, SOCS2, SIRPa and XRCC4 mRNA were calculated. The mean and standard error are shown (p < 0.05).

Article Snippet: The rabbit anti-human primary antibodies (Wuhan Boster Biological Engineering Technology Limited Company) that detect IGFBP5, SOCS1, IL-6 and STAT3(Signal Transducer and Activator of Transcription 3)were incubated with membranes overnight at 4°C.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Microarray, Hybridization

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: The effects of HIF-1alpha on gene expression profiles of NCI-H446 human small cell lung cancer cells

doi: 10.1186/1756-9966-28-150

Figure Lengend Snippet: Western blot analysis of regulation of protein expression by HIF-1alpha in NCI-H446 cells . According to different treatments, all the cells were divided into four groups: control group (the cells cultured under normoxic conditions of 20% O2), Ad5-HIF-1alpha transfection group, hypoxia group (the cells cultured under normoxic conditions of 1% O2) and Ad5-siHIF-1alpha transfection group (after transfection, the cells were cultured under normoxic conditions of 1% O2). (A) Western blot analysis for IGFBP5 protein expressed by the cells of four groups. (B) Western blot analysis for SOCS1 protein expressed by the cells of four groups. (C) Densitometric analysis of the IGFBP5 and SOCS1 bands compared to the corresponding β-actin bands (*p < 0.05 expression of IGFBP5 or SOCS1 protein in Ad5-HIF-1alpha group vs. control group; ** p < 0.05 expression of IGFBP5 or SOCS1 protein in hypoxia group vs. control group; *** p < 0.05 expression of IGFBP5 or SOCS1 protein in Ad5-siHIF-1alpha group vs. control group). (D) Western blot analysis for IL-6 protein expressed by the cells of four groups. (E) Western blot analysis for STAT3 protein expressed by the cells of four groups. (F) Densitometric analysis of the IL-6 and STAT3 bands compared to the corresponding β-actin bands (*p < 0.05 expression of IL-6 or STAT3 protein in Ad5-HIF-1alpha group vs. Ad5-siHIF-1alpha group group.)

Article Snippet: The rabbit anti-human primary antibodies (Wuhan Boster Biological Engineering Technology Limited Company) that detect IGFBP5, SOCS1, IL-6 and STAT3(Signal Transducer and Activator of Transcription 3)were incubated with membranes overnight at 4°C.

Techniques: Western Blot, Expressing, Cell Culture, Transfection

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Expressions of IGFBP-5, cFLIP in cervical intraepithelial neoplasia, cervical carcinoma and their clinical significances: a molecular pathology

doi: 10.1186/1756-9966-28-70

Figure Lengend Snippet: The relationship between expression of IGFBP-5 and cFLIP and clinicopathological parameters in CC

Article Snippet: Primary antibodies used in this study include IGFBP-5 rabbit anti-human polyclonal antibody (Boster Co., Ltd., Wuhan) and cFLIP rabbit anti-human polyclonal antibody (American Neomarker Co.).

Techniques: Expressing

Journal: Acta Neuropathologica

Article Title: Dysregulated IGFBP5 expression causes axon degeneration and motoneuron loss in diabetic neuropathy

doi: 10.1007/s00401-015-1446-8

Figure Lengend Snippet: IGFBP5 protein levels are significantly up-regulated in biopsies of DNP patients. a Western blots of IGFBP5 protein levels in human sural nerve from control (C) individuals (3 samples shown from a total of 5 analyzed), patients (P) with DNP, CIDP, DNP+CIDP (3 samples shown from 4 in total) and other neuropathies (ONP; axonal neuropathy in motor neuron disease (O1, O3), vitamin B12 deficiency (O2). b IGFBP5 protein levels were specifically up-regulated in patients with DNP. Biopsies of patients with DNP showed on average a 5-fold up-regulation compared to CIDP individuals and 50-fold in comparison to healthy controls. Up-regulation in DNP patients with additional CIDP was not significantly higher than in patients with DNP alone. Other neuropathies (ONP) and patients with CIDP showed no significant change in comparison to controls. All values were normalized to CIDP biopsies. QL: quantitative labeling. c IGFBP5 ( green ) and neurofilament ( red ) distribution in human sural nerves from a control individual and a patient with DNP. Note that IGFBP5 levels were increased in axons and in the extracellular matrix (ECM) ( arrows ) in DNP. Scale bar 5 μm

Article Snippet: The membranes were probed with rabbit anti-IGFBP5 (H-100, 1:5000, Santa Cruz Biotechnology) for human samples and goat anti-IGFBP5 antibody (GT15183, 1:5000, Neuromics) for murine samples in 5 % skim milk, and rabbit anti-phosho-IGF1 receptor beta (3918, 1:2000) in 5 % BSA, anti-IGF1 receptor beta (3027, 1:2000) in 5 % milk, for immunoprecipitation 1:1000 in combination with protein A agarose beads (11719386001, Roche), anti-phospho-Akt (9271, 1:2000 in 5 % BSA) and anti-Akt (9272, 1:2000 in 5 % milk, all from Cell Signaling Technology) for mouse extracts, in blocking buffer for 1 h. The appropriate HRP-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, Inc) was used and visualized using enhanced chemiluminescence (GE Healthcare, Lifesciences).

Techniques: Western Blot, Labeling

Journal: Acta Neuropathologica

Article Title: Dysregulated IGFBP5 expression causes axon degeneration and motoneuron loss in diabetic neuropathy

doi: 10.1007/s00401-015-1446-8

Figure Lengend Snippet: IGFBP5 decreases the survival promoting downstream effect of IGF1 on isolated wild-type motoneurons. a IGFBP5 (BP5) reduced IGF1-mediated survival effects from 43 to 20 % on wild-type motoneurons after 7 days in culture, while IGFBP5 did not inhibit BDNF-mediated survival. b Downstream activation of IGF1 signaling pathway in wild-type motoneurons after stimulation with different IGF1 concentrations. Wild-type motoneuron cell cultures were grown for 5 days with 5 ng/ml BDNF, starved overnight and pulsed for 20 min with indicated IGF1 concentrations. Western blot analyses revealed the strongest phosphorylation of IGF1R ( b , c ) and AKT ( b , d ) with 20 ng/ml IGF1. e Downstream activation of IGF1 signaling pathway in wild-type DRGs after stimulation with IGF1. Wild-type DRG cell cultures were grown for 20 h with 10 ng/ml NGF, starved for 4 h and pulsed for 20 min with indicated factors. Western blot analyses revealed the strongest phosphorylation of IGF1R ( e , f ) and AKT ( e , g ) with 20 ng/ml IGF1

Article Snippet: The membranes were probed with rabbit anti-IGFBP5 (H-100, 1:5000, Santa Cruz Biotechnology) for human samples and goat anti-IGFBP5 antibody (GT15183, 1:5000, Neuromics) for murine samples in 5 % skim milk, and rabbit anti-phosho-IGF1 receptor beta (3918, 1:2000) in 5 % BSA, anti-IGF1 receptor beta (3027, 1:2000) in 5 % milk, for immunoprecipitation 1:1000 in combination with protein A agarose beads (11719386001, Roche), anti-phospho-Akt (9271, 1:2000 in 5 % BSA) and anti-Akt (9272, 1:2000 in 5 % milk, all from Cell Signaling Technology) for mouse extracts, in blocking buffer for 1 h. The appropriate HRP-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, Inc) was used and visualized using enhanced chemiluminescence (GE Healthcare, Lifesciences).

Techniques: Isolation, Activation Assay, Western Blot

Journal: Acta Neuropathologica

Article Title: Dysregulated IGFBP5 expression causes axon degeneration and motoneuron loss in diabetic neuropathy

doi: 10.1007/s00401-015-1446-8

Figure Lengend Snippet: Igfbp5 tg + motoneurons show decreased survival when cultured with IGF1. a qRT-PCR revealed a 1.8-fold increase of the Igfbp5 mRNA level in the spinal cord of 6-month-old Igfbp5 tg + mice ( Bp5 tg +). Scored values were obtained by normalizing to β-actin mRNA levels. A.U. arbitrary units. b Igfbp5 protein elevation was detected in sciatic nerve, but not in spinal cord and brain of 6-month-old Igfbp5 tg + mice compared to controls. Actin was used as loading control. c Quantification of Igfbp5 protein levels. QL quantitative labeling. d Motoneurons stained for Tau ( red ) and Igfbp5 ( green ) 7 days after plating. Igfbp5 was increased in the soma and neurites of Igfbp5 tg + motoneurons. Igfbp5 appeared associated with the cell membrane. Scale bar 5 μm. e The positive effect of IGF1 on motoneuron survival is reduced by 37 % in Igfbp5 tg + motoneurons. Motoneuron survival was unchanged in the presence of BDNF

Article Snippet: The membranes were probed with rabbit anti-IGFBP5 (H-100, 1:5000, Santa Cruz Biotechnology) for human samples and goat anti-IGFBP5 antibody (GT15183, 1:5000, Neuromics) for murine samples in 5 % skim milk, and rabbit anti-phosho-IGF1 receptor beta (3918, 1:2000) in 5 % BSA, anti-IGF1 receptor beta (3027, 1:2000) in 5 % milk, for immunoprecipitation 1:1000 in combination with protein A agarose beads (11719386001, Roche), anti-phospho-Akt (9271, 1:2000 in 5 % BSA) and anti-Akt (9272, 1:2000 in 5 % milk, all from Cell Signaling Technology) for mouse extracts, in blocking buffer for 1 h. The appropriate HRP-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, Inc) was used and visualized using enhanced chemiluminescence (GE Healthcare, Lifesciences).

Techniques: Cell Culture, Quantitative RT-PCR, Labeling, Staining

Journal: Acta Neuropathologica

Article Title: Dysregulated IGFBP5 expression causes axon degeneration and motoneuron loss in diabetic neuropathy

doi: 10.1007/s00401-015-1446-8

Figure Lengend Snippet: Reduced phosphorylation of IGF1 receptor in Igfp5 tg + motoneurons leads to reduced survival and axon outgrowth. a Dose–response curve for IGF1-mediated motoneuron survival. Wild-type and Igfp5 tg + motoneurons (7 DIV) were treated with different IGF1 concentrations. b Reduced phosphorylation of IGF1 receptor and AKT in Igfbp5 tg + motoneurons compared to wild-type motoneurons. c Quantification of pIGF1R protein levels in motoneurons. d Quantification of pAKT protein levels in motoneurons. e Images of cultured wild-type motoneurons stained against Tau to identify axons. Scale bar 100 μm. f Igfbp5 (BP5) inhibited IGF1-induced axon outgrowth in wild-type motoneurons. Igfbp5 tg + motoneurons showed shorter axons than wild-type axons when cultured with IGF1. Axon growth stimulated by BDNF was not affected in Igfbp5 tg + motoneurons

Article Snippet: The membranes were probed with rabbit anti-IGFBP5 (H-100, 1:5000, Santa Cruz Biotechnology) for human samples and goat anti-IGFBP5 antibody (GT15183, 1:5000, Neuromics) for murine samples in 5 % skim milk, and rabbit anti-phosho-IGF1 receptor beta (3918, 1:2000) in 5 % BSA, anti-IGF1 receptor beta (3027, 1:2000) in 5 % milk, for immunoprecipitation 1:1000 in combination with protein A agarose beads (11719386001, Roche), anti-phospho-Akt (9271, 1:2000 in 5 % BSA) and anti-Akt (9272, 1:2000 in 5 % milk, all from Cell Signaling Technology) for mouse extracts, in blocking buffer for 1 h. The appropriate HRP-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, Inc) was used and visualized using enhanced chemiluminescence (GE Healthcare, Lifesciences).

Techniques: Cell Culture, Staining

Journal: Acta Neuropathologica

Article Title: Dysregulated IGFBP5 expression causes axon degeneration and motoneuron loss in diabetic neuropathy

doi: 10.1007/s00401-015-1446-8

Figure Lengend Snippet: IGFBP5-overexpressing mice show decreased activation of IGF1 receptor that corresponds to motoneuron degeneration and myelination defects. a , b Immunoprecipitation of the IGF1 receptor from spinal cord ( left panel ) and sciatic nerve ( right panel ) extracts of 4-day-old mice and subsequent analysis of phosphorylation showed decreased activation levels in Igfbp5 tg + mice compared to controls. SN supernatant, IP immunoprecipitation. c Localization of Igfbp5 in cross sections of sciatic nerves. Igfbp5 was increased in axons and extracellular matrix of Igfbp5 tg + mice. Arrows depict extracellular matrix staining. Scale bar 5 µm. d Representative photomicrograph of myelinated fibers in semithin cross sections of the sciatic nerve of 6-month-old wild-type and Igfbp5 tg + animals. Scale bar 10 µm. e 5- to 6-month-old Igfbp5 tg + mice showed a 14 % loss of sciatic nerve fibers. f The frequency of fibers with a circumference between 20–25 μm was decreased by 4.5 % in the sciatic nerve of 6-month-old Igfbp5 tg + mice. g The M-ratio was significantly reduced by 20 % in 6-month-old Igfbp5 tg + mice

Article Snippet: The membranes were probed with rabbit anti-IGFBP5 (H-100, 1:5000, Santa Cruz Biotechnology) for human samples and goat anti-IGFBP5 antibody (GT15183, 1:5000, Neuromics) for murine samples in 5 % skim milk, and rabbit anti-phosho-IGF1 receptor beta (3918, 1:2000) in 5 % BSA, anti-IGF1 receptor beta (3027, 1:2000) in 5 % milk, for immunoprecipitation 1:1000 in combination with protein A agarose beads (11719386001, Roche), anti-phospho-Akt (9271, 1:2000 in 5 % BSA) and anti-Akt (9272, 1:2000 in 5 % milk, all from Cell Signaling Technology) for mouse extracts, in blocking buffer for 1 h. The appropriate HRP-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, Inc) was used and visualized using enhanced chemiluminescence (GE Healthcare, Lifesciences).

Techniques: Activation Assay, Immunoprecipitation, Staining

Journal: Acta Neuropathologica

Article Title: Dysregulated IGFBP5 expression causes axon degeneration and motoneuron loss in diabetic neuropathy

doi: 10.1007/s00401-015-1446-8

Figure Lengend Snippet: Nerve fiber degeneration and spinal motoneuron loss are partly reflected in electrophysiological alterations in motor nerves of Igfbp5 transgenic mice. a Light micrographs of 6-month-old wild-type, Igfbp5 tg + and cIgf1r ko phrenic nerve semithin sections stained with azur–methylene blue. Scale bar 20 µm. b , c Sciatic nerve cross sections of wild-type and Igfbp5 tg + mice. No degenerating fibers were detectable in control tissue and small fibers appeared normal in Igfbp5 tg + mice. Larger fibers of 6-month-old wild-type and Igfbp5 tg + mice ( arrows ) showed signs of degeneration. Scale bar 10 μm. d Nissl-stained paraffin sections of the lumbar spinal cord showed degenerating motoneurons in Igfbp5 tg + mice compared to wild-type animals. Scale bar 50 μm. e 3-week-old Igfbp5 tg + mice showed a 15 % loss of phrenic nerve axons. f , g In 5- to 6-month-old Igfbp5 tg + mice, the number of facial motoneurons was reduced by 17 %, and the number of lumbar spinal motoneurons by 20 %. h 9-Month-old Igfbp5 tg + mice showed reduced grip strength compared with wild-type animals. i Motor nerve conduction velocity (M-NCV) was reduced significantly in Igfbp5 tg + mice by 18 % in sciatic nerve. j Distal compound muscle action potential (CMAP) amplitudes were not altered in Igfbp5 tg + and control animals. k Compound sensory–motor nerve conduction velocity (cSNCV) was not altered in Igfbp5 tg + mice

Article Snippet: The membranes were probed with rabbit anti-IGFBP5 (H-100, 1:5000, Santa Cruz Biotechnology) for human samples and goat anti-IGFBP5 antibody (GT15183, 1:5000, Neuromics) for murine samples in 5 % skim milk, and rabbit anti-phosho-IGF1 receptor beta (3918, 1:2000) in 5 % BSA, anti-IGF1 receptor beta (3027, 1:2000) in 5 % milk, for immunoprecipitation 1:1000 in combination with protein A agarose beads (11719386001, Roche), anti-phospho-Akt (9271, 1:2000 in 5 % BSA) and anti-Akt (9272, 1:2000 in 5 % milk, all from Cell Signaling Technology) for mouse extracts, in blocking buffer for 1 h. The appropriate HRP-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, Inc) was used and visualized using enhanced chemiluminescence (GE Healthcare, Lifesciences).

Techniques: Transgenic Assay, Staining