Journal: Journal of Cardiovascular Development and Disease

Article Title: Angiotensin II Receptor Blocker Irbesartan Enhanced SIRT1 longevity Signaling Replaces the Mitochondrial Biogenetic Survival Pathway to Attenuate Hypertension-Induced Heart Apoptosis

doi: 10.3390/jcdd9080266

Figure Lengend Snippet: ( A ) The representative protein levels of IGF-II and p -JNK from the left ventricles in the normotensive group (WKY), untreated hypertensive group (SHR), and SHR rats with treated irbesartan (SHR-ARB), as measured by Western blot methods. The α-Tubulin served as an internal control. ( B ) Bars indicate the relative fold changes of protein levels relative to the Control group in IGF-II and p -JNK on α-Tubulin and show mean ± standard deviation (n = 8;8;8). ** p < 0.01: significant differences from the Control group. ## p < 0.01 indicates significant differences between the SHR group and the SHR-ARB group.

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies, including Fas Ligand, Fas, FADD, Bax, cytosolic cytochrome c, active caspase-8, active caspase-9, and active caspase-3, IGF-II, p -JNK, SIRT1, PGC-1, Bcl-2, Bcl-xL, and α-tubulin (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Western Blot, Control, Standard Deviation

Journal: Journal of Cardiovascular Development and Disease

Article Title: Angiotensin II Receptor Blocker Irbesartan Enhanced SIRT1 longevity Signaling Replaces the Mitochondrial Biogenetic Survival Pathway to Attenuate Hypertension-Induced Heart Apoptosis

doi: 10.3390/jcdd9080266

Figure Lengend Snippet: Hypothesized diagram. The proposed schematic diagram from the current study shows that the cardiac Fas/FasL-mediated (Fas Ligand, Fas, FADD, and active caspase-8) and the mitochondria-mediated apoptotic pathways (Bax, cytosolic cytochrome c, Active caspase-9, and Active caspase-3) were activated by hypertension through IGF-II and p -JNK activation. Moreover, the pro-survival protein of SIRT1 appeared to have a compensatory pro-survival response after hypertension. However, these cardiac Fas/FasL-mediated and mitochondria-mediated apoptotic pathways appear to be suppressed or not activated via the enhanced SIRT1/PGC-1α pro-survival pathway (SIRT1, PGC-1, Bcl-2, and Bcl-xL) under hypertension after treated Irbesartan.

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies, including Fas Ligand, Fas, FADD, Bax, cytosolic cytochrome c, active caspase-8, active caspase-9, and active caspase-3, IGF-II, p -JNK, SIRT1, PGC-1, Bcl-2, Bcl-xL, and α-tubulin (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Activation Assay

Journal: Translational Pediatrics

Article Title: Development and external validation of a mitophagy-related diagnostic model for biliary atresia based on cellular infiltration patterns

doi: 10.21037/tp-2025-1-864

Figure Lengend Snippet: Single cell sequencing and immunohistochemistry validate key molecular markers for BA. (A) Representative immunohistochemistry images validating the protein expression of mitophagy-related genes ( IGF2BP3 , NEDD4L , and ALDH2 ) in liver tissues from BA patients and controls (scale bar =100 µm). Ctrl images are from non-tumor liver tissue adjacent to hepatoblastoma specimens. Positive staining appears as brown discoloration and infiltrating immune cells were indicated by red arrows. (B) Differential analysis of single-cell transcription. (a) Single-cell’s cell classification landscape in all samples. (b) The single-cell expression differences of IGF2BP3, NEDD4L, PPIB, and ALDH2 between BA and Ctrl groups. BA, biliary atresia; Ctrl, control; FC, fold change.

Article Snippet: Next, sections were incubated overnight at 4 °C with the following primary antibodies: anti-human ALDH2 (HUABIO, Hangzhou, China; M1509-1, 1:800), NEDD4L (Proteintech, Wuhan, China; 13690-1-AP, 1:800), and IGF2BP3 (Proteintech, Wuhan, China; 14642-1-AP, 1:800).

Techniques: Single Cell, Sequencing, Immunohistochemistry, Expressing, Staining, Control

Journal: Translational Pediatrics

Article Title: Development and external validation of a mitophagy-related diagnostic model for biliary atresia based on cellular infiltration patterns

doi: 10.21037/tp-2025-1-864

Figure Lengend Snippet: The interaction between drugs and IGF2BP3 and NEDD4L proteins. (A-C) The interaction between cyclosporine and IGF2BP3 protein. (D-F) The interaction between cyclosporine and NEDD4L protein. Blue represents drug molecules, with red indicating oxygen atoms within the molecules. Green represents amino acid residues of the protein interacting with the drug molecules. The yellow background indicates the average interaction energy between the protein and drugs (average of 10 simulations).

Article Snippet: Next, sections were incubated overnight at 4 °C with the following primary antibodies: anti-human ALDH2 (HUABIO, Hangzhou, China; M1509-1, 1:800), NEDD4L (Proteintech, Wuhan, China; 13690-1-AP, 1:800), and IGF2BP3 (Proteintech, Wuhan, China; 14642-1-AP, 1:800).

Techniques:

Journal: Translational Pediatrics

Article Title: Development and external validation of a mitophagy-related diagnostic model for biliary atresia based on cellular infiltration patterns

doi: 10.21037/tp-2025-1-864

Figure Lengend Snippet: Potential schematic representation of the BA-CIMDGM genes in the pathogenesis of BA. Upregulated IGF2BP3 in T cells and NEDD4L in macrophages leads to impaired mitophagy and promotes immune activation and cytokines released. And the downregulation of ALDH2 in hepatocytes compromises the mitochondrial antioxidant defense and excessive ROS production, synergistically driving biliary epithelial cell injury and progressive liver fibrosis. BA, biliary atresia; BA-CIMDGM, Biliary Atresia Cell Infiltration Mitophagy Diagnostic Gene Model; ROS, reactive oxygen species.

Article Snippet: Next, sections were incubated overnight at 4 °C with the following primary antibodies: anti-human ALDH2 (HUABIO, Hangzhou, China; M1509-1, 1:800), NEDD4L (Proteintech, Wuhan, China; 13690-1-AP, 1:800), and IGF2BP3 (Proteintech, Wuhan, China; 14642-1-AP, 1:800).

Techniques: Activation Assay, Diagnostic Assay

Journal: Nature

Article Title: Designed endocytosis-inducing proteins degrade targets and amplify signals.

doi: 10.1038/s41586-024-07948-2

Figure Lengend Snippet: Fig. 1 | Design strategies for endocytosis-triggering EndoTags. a, Schema of designed endocytosis mechanisms. Top, design of binding to constitutively cycling receptors at sites that do not overlap with binding sites for natural ligands to avoid competition. Middle, design of binders that trigger endocytosis by eliciting conformational changes in the receptor. The EndoTag binds at two distinct epitopes on the target and actively triggers the conformational change. Bottom, designed endocytosis via receptor clustering. The multivalent EndoTag clusters multiple copies of the target receptor and induces endocytosis. b, Design strategy for sortilin48 and TfR49 EndoTags. c, Cellular uptake of 100 nM AF647-labelled Sort_EndoTags, TfR-EndoTags or LHDB22 scaffold control for 2 h in U-251MG cells. Data were normalized to the 100 nM AF647-labelled LHDB group (no endocytosis). MFI, mean fluorescence intensity. d, Confocal imaging of Sort_EndoTag (red) and lysosomal marker (green, AF488-labelled LysoTracker) after 24 h incubation in U-251MG cells. e, Design strategy for

Article Snippet: Anti-LAMP2A antibody (Abcam ab18528), goat anti-rabbit- IgG Alexa FluorTM 488 secondary antibody (Thermo Fisher A-11034), anti PD-L1 (sc-518027), anti beta-actin (sc-47778), goat anti-mouse IgG H&L (HRP) (Abcam, ab205719); 800CW goat-anti-mouse or goatanti-rabbit (LI-COR 926-32211), rabbit anti-CTLA4 E1V6T Cell Signaling Technologies (96399) Validation Alexa Fluor® 647 anti-human EGFR Antibody (Biolegend, 352918): Verified Reactivity to Human, FC - Quality tested PE anti-human CD222 (IGF2R) Recombinant Antibody (Biolegend, 364204): Verified Reactivity to Human, ICFC, FC - Quality tested EGFR Monoclonal Antibody (clone 199.12, Invitrogen, AHR5072): Target Species: Human, applications for Immunocytochemistry, Immunofluorescence, Immunoprecipitation, Western Blot IGF2R Antibody Alexa647 (clone 2G11, Novus, NB300-514AF647): Reactivity Hu, Mu, Rt, Bv, Pm; Applications WB, ELISA, Flow, ICC/IF, IHC, IP, CyTOF-ready Human PD-L1 Alexa Fluor® 647-conjugated Antibody (Rndsystems, FAB1562R): Species Reactivity Human, Detects human PD-L1/B7H1 in direct ELISAs.

Techniques: Binding Assay, Control, Fluorescence, Imaging, Marker, Incubation

Journal: Nature

Article Title: Designed endocytosis-inducing proteins degrade targets and amplify signals.

doi: 10.1038/s41586-024-07948-2

Figure Lengend Snippet: Fig. 3 | Clearance of soluble proteins by IGF2R pLYTACs. a, Schema for the use of soluble pLYTACs with IGF_EndoTags. b, Cellular uptake of LHDB–AF647 via LHDA–IGF_EndoTags in Jurkat cells. Cells were incubated with 33 nM LHDB– AF647 with or without 1 μM LHDA–IGF_EndoTags for 24 h, washed twice with cold PBS and analysed by flow cytometry. c, Remaining supernatant LHDB– AF647 levels in Jurkat cells. Jurkat cells were incubated with 100 nM LHDB– AF647 with or without 500 nM LHDA–IGF_EndoTags. At timepoints 24 h and 48 h, the cells were pelleted down, and IgG in the supernatant was quantified using a Neo2 plate reader. IgG level was normalized to the IgG-alone control group. d, Cellular uptake of IgG–AF647 via protein G–IGF_EndoTags in K562 cells. Cells were incubated with 33 nM IgG–AF647 with or without 1 μM protein G–IGF_EndoTag3 for 24 h, washed twice with cold PBS and analysed by flow cytometry. The fold change in MFI was calculated by normalizing to the

Article Snippet: Anti-LAMP2A antibody (Abcam ab18528), goat anti-rabbit- IgG Alexa FluorTM 488 secondary antibody (Thermo Fisher A-11034), anti PD-L1 (sc-518027), anti beta-actin (sc-47778), goat anti-mouse IgG H&L (HRP) (Abcam, ab205719); 800CW goat-anti-mouse or goatanti-rabbit (LI-COR 926-32211), rabbit anti-CTLA4 E1V6T Cell Signaling Technologies (96399) Validation Alexa Fluor® 647 anti-human EGFR Antibody (Biolegend, 352918): Verified Reactivity to Human, FC - Quality tested PE anti-human CD222 (IGF2R) Recombinant Antibody (Biolegend, 364204): Verified Reactivity to Human, ICFC, FC - Quality tested EGFR Monoclonal Antibody (clone 199.12, Invitrogen, AHR5072): Target Species: Human, applications for Immunocytochemistry, Immunofluorescence, Immunoprecipitation, Western Blot IGF2R Antibody Alexa647 (clone 2G11, Novus, NB300-514AF647): Reactivity Hu, Mu, Rt, Bv, Pm; Applications WB, ELISA, Flow, ICC/IF, IHC, IP, CyTOF-ready Human PD-L1 Alexa Fluor® 647-conjugated Antibody (Rndsystems, FAB1562R): Species Reactivity Human, Detects human PD-L1/B7H1 in direct ELISAs.

Techniques: Incubation, Flow Cytometry, Control

Journal: Molecular cancer research : MCR

Article Title: Intrinsic Resistance to Cixutumumab is Conferred by Distinct Isoforms of the Insulin Receptor

doi: 10.1158/1541-7786.MCR-15-0279

Figure Lengend Snippet: Insulin-like growth factor II (IGF-II) mediates resistance to cixutumumab in tumor cells over-expressing IR-B. Clonogenic assay in A549 cells that overexpress IR-B (A) in the presence or absence of cixutumumab and/or anti-IGF-II mAb. Represented are mean values +/− SEM, expressed as percentage of colony formation relative to control treated cells. Statistical significance was determined by one-way ANOVA followed by Tukey’s posthoc analysis. Statistically significant difference vs control treated cells (a), cixutumumab-treated cells (b), anti-IGF-II treated cells (c). (B, C) Western blot analysis of signal transduction in A549-IR-B cells treated with IGF-II (25 nM), in the presence or absence of cixutumumab and linsitinib. Serum-starved cells were pretreated with cixutumumab (100 nM) or linsitinib (0.01, 0.1 or 1 uM) for 15 min and 2 h, respectively, followed by incubation with rhIGF-II (25 nM) for 10 minutes. Proteins (15–20 ug) extracted from cultured cells were size-fractionated by SDS-PAGE and immunoblotted with anti-phospho-IRβY1150/51/IGF-IRβY1135/36, anti-phospho-AktS473 and anti-phospho-p42/p44 MAPKT202/Y204 antibodies. Total level of proteins was demonstrated by immunoblotting with antibodies directed against total Akt and p42/p44 MAPK. (D) MCF-7 and A549 cells with or without IR-A or IRB overexpression were treated with increasing concentrations of linsitinib (0.00015-10 uM) for 72 h, and tumor cell viability was quantified by CellTiter-Glo assay. The results are expressed as % of inhibition of tumor cell viability.

Article Snippet: The following antibodies were purchased from commercial sources as indicated: mouse monoclonal antibodies against Akt (#2920) and p42/p44 MAPK (Erk1/2) (#9107), rabbit monoclonal antibodies against phospho-Akt S473 (#4060) and phospho-IGF-IR beta Y1135/1136 / Insulin Receptor beta Y1150/1151 (#3024), rabbit polyclonal antibody against phospho-p42/p44 MAPK T202/Y204 (Erk1/2) (#9101) (Cell Signalling, Beverly, MA, USA); mouse monoclonal antibody against IGF-IR (#MS-641-P) and Insulin Receptor (#MS-632-P) (Thermo Fisher Scientific, Fremont, CA, USA); rabbit polyclonal Insulin Receptor (#sc-711) and (#sc-7953) (Santa Cruz Biotechnology, Santa Cruz, CA, USA); mouse monoclonal antibody against IGF-II (MAB292) and goat F(ab') 2 anti-mouse IgG-phycoerythrin (#F0102B) (R&D Systems, Minneapolis, MN, USA); goat polyclonal anti-mouse IRDye 680 conjugated (#926–32220) and anti-rabbit IRDye 800 conjugated (#926–32211) (LI-COR Biosciences, Lincoln, Nebraska, USA).

Techniques: Expressing, Clonogenic Assay, Control, Western Blot, Transduction, Incubation, Cell Culture, SDS Page, Over Expression, Glo Assay, Inhibition

Journal: The Journal of Neuroscience

Article Title: Selective Expression of Insulin-Like Growth Factor II in the Songbird Brain

doi: 10.1523/JNEUROSCI.17-18-06974.1997

Figure Lengend Snippet: Diagram highlighting the song nuclei that express IGF-II mRNA (dark gray) and outlining their relationships (thick black arrows) as well as relationships among other song nuclei (thin black andwhite arrows). The two identified populations of projection neurons of HVC, those that project to RA (black) and those that project to area X (white), are schematically indicated. The direct descending motor pathway includes HVC, theRA, and the tracheosyringeal portion of the hypoglossal nucleus (nXIIts). The anterior forebrain pathway is necessary for song acquisition and connects (white arrows) HVC, area X the medial portion of the dorsolateral thalamic nucleus (DLM), and the lateral magnocellular nucleus of the anterior neostriatum (lMAN), which continues to RA and feeds back to area X (Okuhata and Saito, 1987; Bottjer et al., 1989;Nixdorf-Bergweiler et al., 1995; Vates and Nottebohm, 1995). Neurons inRA give rise to two feedback loops. The first one connects RA with the posterior portion of the dorsomedial thalamic nucleus (DMP), which in turn projects to themMAN that synapses with HVC (Vates et al., 1997). The second feedback loop from RA projects to the thalamic nucleus DLM, which in turn projects tolMAN (Wild, 1993; Vates et al., 1997).

Article Snippet: IGF-II-like immunoreactivity was detected using a polyclonal antibody against human IGF-II (Gropep, Australia).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Selective Expression of Insulin-Like Growth Factor II in the Songbird Brain

doi: 10.1523/JNEUROSCI.17-18-06974.1997

Figure Lengend Snippet: Complete nucleotide sequence of the zebra finch cDNA clone pIGF-II-ZF and deduced primary structure of the corresponding IGF-II prepropeptide. Translation start site, stop codon (star), and the mature peptide are in bold letters. The ORF of 561 nucleotides was flanked by 323 nucleotides of 5′-UTR and 478 nucleotides of 3′-UTR. Nucleotide positions are given on the right, starting with the ATG site. The numbering of the amino acid residues is given in italics and starts at the N terminal of the mature IGF-II peptide.

Article Snippet: IGF-II-like immunoreactivity was detected using a polyclonal antibody against human IGF-II (Gropep, Australia).

Techniques: Sequencing

Journal: The Journal of Neuroscience

Article Title: Selective Expression of Insulin-Like Growth Factor II in the Songbird Brain

doi: 10.1523/JNEUROSCI.17-18-06974.1997

Figure Lengend Snippet: Northern analysis of zebra finch total and poly(A+)-selected RNA. A, Samples of total RNA from the head region of embryonic day 14 (E14) and E16 embryos and from the forebrain of 45-d-old (D45) and D120birds were hybridized with the IGF-II antisense riboprobe, revealing multiple transcripts ranging in size between 0.9 and 4.0 kb.B, The sense probe showed no hybridization to RNA samples from embryonic head (E16) or adult forebrain (D120). C, After poly(A+) selection of embryonic (pooled E6–E14) RNA and hybridization with the antisense probe under identical conditions, a 4.0 kb mRNA was recognized. Ribosomal RNA locations28S and 18S were determined from the ethidium bromide-stained gel.

Article Snippet: IGF-II-like immunoreactivity was detected using a polyclonal antibody against human IGF-II (Gropep, Australia).

Techniques: Northern Blot, Hybridization, Selection, Staining

Journal: The Journal of Neuroscience

Article Title: Selective Expression of Insulin-Like Growth Factor II in the Songbird Brain

doi: 10.1523/JNEUROSCI.17-18-06974.1997

Figure Lengend Snippet: IGF-II mRNA expression in song nuclei of a 2.5-year-old canary. A, In a dark-field photomicrograph of a parasagittal section ∼2 mm from the midline, a strong IGF-II mRNA ISH signal is seen as white silver grains overlying HVC and to a lesser extent over RA. C, Approximately 500 μm from the midline, mMAN shows a strong IGF-II mRNA signal. Acrescent-shaped area with selective expression is visible in the most caudal reaches of Ncmas well as a layer of cells in the parahippocampus. Leptomeninges have been stripped off the brain during dissection except around the olfactory bulb, in which they show the characteristically strong IGF-II expression. B, D, Anatomical diagrams of sections shown to the left. A, Archistriatum; APH, area parahippocampalis;BO, olfactory bulb; Cb, cerebellum;Ch.O., chiasma opticum; FA, frontoarchistriatal tract; HA, hyperstriatum accessorium; HP, hippocampus; HV, hyperstriatum ventrale; LAD, lamina archistriatalis dorsalis; LFM, lamina frontalis suprema;LH, lamina hyperstriatica; LMD, lamina medularis dorsalis; LPO, lobus parolfactorius;N, neostriatum; Ncm, caudomedial neostriatum. Scale bars, 1 mm.

Article Snippet: IGF-II-like immunoreactivity was detected using a polyclonal antibody against human IGF-II (Gropep, Australia).

Techniques: Expressing, Dissection, Olfactory

Journal: The Journal of Neuroscience

Article Title: Selective Expression of Insulin-Like Growth Factor II in the Songbird Brain

doi: 10.1523/JNEUROSCI.17-18-06974.1997

Figure Lengend Snippet: Cellular expression of IGF-II mRNA in canary (A, C, E,G) and zebra finch (H) song nuclei and negative controls (B, D,F) hybridized with the sense probe.A, HVC shows a uniform distribution of IGF-II-labeled cells and a homogeneous signal intensity throughout the area.Arrowheads (A–D) indicate the position of the lateral ventricle. C, Higher magnification reveals that not all cells in HVC are labeled and that the ventricular zone is negative. E, Within the cell clusters typical for HVC, IGF-II mRNA is often expressed in a subset of HVC cells (black arrows) surrounded by IGF-II negative cells. Nomarski optics highlight cell nuclei as darker, round profiles surrounded by a lighter-appearing cytoplasmic region, the boundaries of which cannot be seen. The grain distribution of theupper IGF-II-positive cell indicates labeled mRNA both in the cytoplasm overlying the nucleus and in the cytoplasm extending beyond the nucleus to the top right. B,D, F, In control sections hybridized with the sense strand, adjacent to those sections shown in A,C, and E, respectively, label is not different from background. G, IGF-II is strongly expressed in adult canary mMAN. H, IGF-II-positive cells are uniformly distributed in adult zebra finch RA, although this intensity of label was rare. White arrows indicate the anatomical outline of the RA; white arrowheads point to the IGF-II-positive meninges. All photomicrographs were obtained from parasagittal sections (dorsal is at the top, and rostral is to the right) under dark-field (A,B, G, H) or Nomarski illumination (C–F). Scale bars: A, B, 500 μm;C, D, 50 μm; E,F, 10 μm; G, H, 250 μm.

Article Snippet: IGF-II-like immunoreactivity was detected using a polyclonal antibody against human IGF-II (Gropep, Australia).

Techniques: Expressing, Labeling, Control

Journal: The Journal of Neuroscience

Article Title: Selective Expression of Insulin-Like Growth Factor II in the Songbird Brain

doi: 10.1523/JNEUROSCI.17-18-06974.1997

Figure Lengend Snippet: Conspicuous IGF-II mRNA expression was found in the hippocampus (A), cerebellar Purkinje cells (C), brainstem (E), and choroid plexus (A, G). B,D, F, H, Negative controls hybridized with the sense probe show signals not different from background. A, A low-magnification dark-field photomicrograph shows a streak of IGF-II-positive cells (white arrowheads) in the hippocampus of an adult canary. The IGF-II-positive meninges are indicated with a black arrowhead. Note also that the choroid plexus (white arrow) is strongly labeled. C, Cerebellar Purkinje cells, shown here in an adult zebra finch, were strikingly labeled (arrows). Often deep cerebellar nuclei were also positive. E, Several pontomesencephalic nuclei showed IGF-II mRNA expression, among them AVT (arrow) shown here in a dark-field photomicrograph of an adult canary section.G, A high-power light-field photomicrograph shows that IGF-II mRNA is primarily expressed in the epithelium (arrows) of the choroid plexus, shown here in an adult zebra finch. All sections are cut in the parasagittal plane ∼500 μm from the midline; dorsal is at the top, and caudal is to the left. Scale bars:A–F, 1 mm; G,H, 10 μm.

Article Snippet: IGF-II-like immunoreactivity was detected using a polyclonal antibody against human IGF-II (Gropep, Australia).

Techniques: Expressing, Labeling

Journal: The Journal of Neuroscience

Article Title: Selective Expression of Insulin-Like Growth Factor II in the Songbird Brain

doi: 10.1523/JNEUROSCI.17-18-06974.1997

Figure Lengend Snippet: Developmental regulation of zebra finch HVC-specific IGF-II mRNA expression, quantified by image analysis of ISH signal. IGF-II levels in HVC were highest in group 2 (mean age of 68 d). Both younger juveniles (group 1, mean age of 42 d) and adults (group 3, between 93 d and 4.7 years old) had significantly lower levels (group 1 vs group 2, p = 0.0001; group 2 vs group 3, p < 0.05; group 1 vs group 3,p > 0.05). HVC-specific expression was obtained by subtracting the grain density values measured in an area ventral to HVC from those measured inside HVC. The group differences are caused by changing mRNA expression within HVC because expression patterns in the area below HVC did not vary among groups. The differences between groups are expressed in optical density units, and error bars indicate SEM. Significances were calculated using ANOVA and Scheffé’sF tests and post hoc Student’st test comparisons (ANOVA, n = 21;F = 14; p = 0.0002).

Article Snippet: IGF-II-like immunoreactivity was detected using a polyclonal antibody against human IGF-II (Gropep, Australia).

Techniques: Expressing

Journal: The Journal of Neuroscience

Article Title: Selective Expression of Insulin-Like Growth Factor II in the Songbird Brain

doi: 10.1523/JNEUROSCI.17-18-06974.1997

Figure Lengend Snippet: Comparison of seasonal variation of (A) neuron addition and (B) IGF-II mRNA expression in HVC of male canaries between 1 and 2 years of age, sampled each month throughout a 12 month period. A, Neuron addition to the HVC was not the same during all months (ANOVA,F = 7.3; p = 0.0001); Reproduced with permission from Kirn et al. (1994). B, HVC-specific IGF-II mRNA was present at varying levels throughout the year, but differences were not statistically significant (ANOVA,n = 45; F = 1.2;p = 0.3448). However, a comparison ofA and B shows that significant peaks in neuron addition coincide with (relative to surrounding months) high IGF-II mRNA levels (October and March, darkly shaded bars). Moreover, month-to-month changes are in the same direction for both neuron addition and IGF-II levels, with the exception of April to May. C, Monthly changes in neuron addition and IGF-II levels covary significantly (n= 11; r = 0.595; p < 0.05). Monthly changes in neuron addition were calculated from the original data generated by Kirn et al. (1994) and were used to plot A. Monthly changes in IGF-II expression levels were calculated from the data shown in B. Each point on the scatterplot (C) represents one monthly transition, e.g., June-to-July, July-to-August, etc. Theletters on the x-axis forA and B are the first letter for each month of the year, starting on the left with June.

Article Snippet: IGF-II-like immunoreactivity was detected using a polyclonal antibody against human IGF-II (Gropep, Australia).

Techniques: Comparison, Expressing, Generated

Journal: The Journal of Neuroscience

Article Title: Selective Expression of Insulin-Like Growth Factor II in the Songbird Brain

doi: 10.1523/JNEUROSCI.17-18-06974.1997

Figure Lengend Snippet: IGF-II immunoreactivity in parasagittal sections of adult canary forebrain. A, Immunoreactive material accumulates within HVC but is absent in neurons from the surrounding tissue. B, Preincubation of the antibody with human recombinant IGF-II abolishes specific IGF-II immunoreactivity in HVC. The anatomical outline of HVC is indicated with arrowheads. The hippocampus overlying HVC has detached from the section and partially overlaps with the dorsal edge of HVC. C, At high magnification the IGF-II peptide staining is perinuclear and curvilinear, a pattern suggesting Golgi-associated localization. D, IGF-II immunoreactivity is present in mMAN, indicated by anarrowhead. E, Anatomical identification of mMAN (arrowhead) by means of retrogradely biotinylated dextranamine-labeled neurons, as described in Vates et al. (1997). F, IGF-II antibody also stains a stellate cell type, probably microglia. G, IGF-II-positive stellate cells are much more abundant in LPO than in neostriatum.H, The radial cells typical of the adult songbird brain showed strong IGF-II immunoreactivity in their processes. Awhite arrow indicates the anterior edge of HVC in which IGF-II-positive cells can also be seen. The lateral ventricle overlying HVC is marked by an arrowhead. I, Close-up of a radial fiber with its characteristic thickening.K, Punctate staining in the ventricular zone of the lateral ventricle indicates the presence of IGF-II immunoreactive material. HV, Hyperstriatum ventrale;LPO, lobus parolfactorius; N, neostriatum. Scale bars: A, B,D, E, 500 μm; C,F–I, K, 10 μm.

Article Snippet: IGF-II-like immunoreactivity was detected using a polyclonal antibody against human IGF-II (Gropep, Australia).

Techniques: Recombinant, Staining, Labeling

Journal: The Journal of Neuroscience

Article Title: Selective Expression of Insulin-Like Growth Factor II in the Songbird Brain

doi: 10.1523/JNEUROSCI.17-18-06974.1997

Figure Lengend Snippet: Combined labeling with retrograde neuronal markers, nonradioactive ISH, and ICC in canary brain. A, HVC was photographed under fluorescent light for the identification of area X-projecting neurons, labeled with retrogradely transported rhodamine microspheres (red label, white arrows). B, The same field shown inA under bright-field illumination shows the presence of IGF-II mRNA detected by nonradioactive ISH (black label,black arrows) in the area X-projecting cells identified in A. C, Retrogradely rhodamine (bright red)-labeled RA-projecting neurons (white arrows) do not overlap with, but are adjacent to, nonradioactive IGF-II ISH-labeled neurons (black arrows). D, Bright-field photomicrograph of the same field shown in C focusing on the same ISH-labeled cells dimly visible in C. White arrowspoint to the absence of ISH label in the places in which rhodamine-positive RA-projecting cells are visible in C.E, Combination of retrograde labeling with ICC showed that IGF-II immunoreactivity (black label, black arrowheads) is not present in, but is adjacent to, the area X-projecting neurons (red label, white arrows). F, Double labeling using the IGF-II antibody (black, photographed under bright-field illumination) and retrograde Fluorogold filling of the HVC-to-RA-projecting neurons (white, photographed under UV illumination) indicates that IGF-II immunoreactivity accumulates in these cells (black arrowheads). The majority of IGF-II-positive cells is Fluorogold-labeled, but a few immunopositive cells are not backfilled (white arrowheads)G, Triple-labeled HVC section shows IGF-II-specific immunoreactivity overlying Fluorogold-labeled RA-projecting cells (black arrowheads), whereas rhodamine microsphere-labeled area X-projecting cells (white arrows) are immunonegative. Cells in the focal plane of the photo most clearly show Fluorogold (whitish-blue) and IGF-II immunoreactivity (dark-blue/black). Fluorogold label in cells deeper in the tissue is attenuated by the section thickness and appears dimmer. Section thicknesses:A–D, 10 μm; F, 20 μm;E, G, 40 μm. Scale bars, 10 μm.

Article Snippet: IGF-II-like immunoreactivity was detected using a polyclonal antibody against human IGF-II (Gropep, Australia).

Techniques: Labeling