goat anti igfbp2  (R&D Systems)

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: IGFBP2 regulated genes in BT474 breast cancer cells. a ) Western blot analysis of IGFBP2 in the supernatant of IGFBP2 knockdown clones C5 and C12 and control cells. Lower panel is ponceau stained membrane shown as loading control. b ) Hierarchical cluster of differentially expressed genes in IGFBP2 knockdown BT474 cells versus control cells. Differentially regulated genes were clustered using MeV software. The dendrogram on the left shows different clusters of genes segregated according to the pattern of regulation. Red and green indicate high and low expression of genes respectively. Black indicates no regulation. c ) Validation of selected genes by qPCR. Bar graphs of differentially regulated genes in IGFBP2 Knockdown BT474 clones versus control cells. The graphs represent the fold change over control after normalization with the expression of RPL35a.

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques: Western Blot, Clone Assay, Staining, Software, Expressing

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: List of top 50 differentially regulated genes (p < 0.05) in IGFBP2 positive versus IGFBP2 negative tumors

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques:

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: Differential expression of genes in IGFBP2 positive and IGFBP2 negative tumor samples compared to control tissues. a ) Genes were clustered using MeV software. The dendrogram on the left shows different clusters of genes segregated according to the pattern of regulation. Red and green indicate high and low expression of genes respectively. Black indicates no regulation. b ) Validation of selected genes. Scatter plots of differentially regulated genes in IGFBP2 positive and IGFBP2 negative tumors compared to the expression in normal tissues. Log 2-transformed gene expression ratios obtained from real-time quantitative PCR analysis normalized to TBP expression are plotted. Each dot represents a data derived from one sample.

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques: Expressing, Software, Transformation Assay, Real-time Polymerase Chain Reaction, Derivative Assay

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: List of top 50 common genes differentially regulated between IGFBP2 positive tumors and IGFBP2 knockdown clones

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques:

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: β-catenin and IGFBP2 expression in IGFBP2 knockdown and control cells. IGFBP2 knockdown clones and control cells were plated on coverslips and allowed to grow. 24 h after plating, cells were fixed, permeabilized, and stained for β-catenin and IGFBP2 expression. Expression of β-catenin and IGFBP2 is shown in green and blue, respectively. Nucleus was stained using propidium iodide (PI) as shown in red. Original magnification was 63×.

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques: Expressing, Clone Assay, Staining

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: Regulation of β-catenin by IGFBP2 is IGF1R and FAK dependent. IGFBP2 knockdown clone C12 was transfected with IGFBP2 and 36 h. after transfection cells were fixed and analyzed for β-catenin, and IGFBP2. For inhibitor experiments, 24 h. after transfection cells were treated with IGF1R or FAK inhibitor for 12 h. Cells were fixed and analyzed for β-catenin and IGFBP2 expression. Expression of β-catenin and IGFBP2 is shown in green and blue, respectively. Nucleus was stained using propidium iodide (PI) as shown in red. Original magnification was 63×. V, Vector control; OE, Over expression; Inh, inhibitor.

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques: Transfection, Expressing, Staining, Plasmid Preparation, Over Expression

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: β-catenin and IGFBP2 expression in breast cancer tissues. Representative micrographs showing β-catenin staining in cancer tissues. a β-catenin staining of a section of IDC 3 breast tumors showing cytoplasmic and membrane staining. b represents breast cancer tissue section stained for IGFBP2 showing predominantly cytoplasmic staining.

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques: Expressing, Staining

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: IGFBP2 and β-catenin expression in breast cancer tissues

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques: Expressing

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: List of top 50 differentially regulated genes (p < 0.05) in IGFBP2 knockdown clones

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques: shRNA

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: GSEA summary of pathways associated with genes down regulated upon IGFBP2 knockdown

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques:

Journal: Molecular Cancer

Article Title: Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with β-catenin in breast cancer lymph node metastasis

doi: 10.1186/1476-4598-12-63

Figure Lengend Snippet: GSEA summary of pathways associated with genes up regulated in IGFBP2 positive tumors

Article Snippet: After the overnight incubation, the cells were washed thrice with PBS and incubated with the secondary antibody, 1:1500 dilution of alexa flur 488 (anti-rabbit, for β-catenin) and alexa flur 633 (anti-goat for IGFBP2) (Molecular probes, Invitrogen, USA) in PBS for 1 hour in dark.

Techniques:

Journal: Aging (Albany NY)

Article Title: Intracellular Insulin-like growth factor binding protein 2 (IGFBP2) contributes to the senescence of keratinocytes in psoriasis by stabilizing cytoplasmic p21

doi: 10.18632/aging.103045

Figure Lengend Snippet: Psoriatic keratinocyte cultures display enhanced IGFBP2 expression, together with an altered expression of genes implicated in the regulation and cell cycle arrest. ( A ) Real-time PCR analysis was performed on keratinocyte cultures (at passage P4), obtained from lesional skin of psoriatic patients ( n = 6) (pso KC) and healthy volunteers ( n = 6) (healthy KC). Results are shown as individual values of relative mRNA levels (normalized to β-actin) of IGFBP2, IGFBP3, p16, p21 Cdk1, cyclin A and p57 and means of the two different groups. ( B ) WB analysis was performed on protein lysates from keratinocyte cultures isolated from healthy ( n = 6) and lesional skin ( n = 6) by using anti-IGFBP2, cyclin A, cdk1, -p16 and -p21 Abs. β-actin was used as loading control. Bands relative to IGFBP2 were showed at two different exposure times (High exp. 1 min; low exp., 30 seconds). Graphs represent the individual values and the means of the densitometric intensity (D.I.) of each band. ( A , B ), * p ≤ 0.05, as calculated by the Mann–Whitney U test.

Article Snippet: For the specific neutralization of IGFBP2 bioactivity in keratinocyte supernatants, the neutralizing goat polyclonal anti-human IGFBP2 Ab (AF674) or goat IgG control (AB-108-C) from R&D Systems was used.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Isolation, Control, MANN-WHITNEY

Journal: Aging (Albany NY)

Article Title: Intracellular Insulin-like growth factor binding protein 2 (IGFBP2) contributes to the senescence of keratinocytes in psoriasis by stabilizing cytoplasmic p21

doi: 10.18632/aging.103045

Figure Lengend Snippet: IGFBP2 is enhanced in the suprabasal layers of lesional psoriatic skin, and parallels p16 and p21 senescence markers. ( A ) IHC analysis of IGFBP2, p21 and phosphorylated p21 (p-p21, red-brown stained), as well as of p16 (red-stained), was performed on paraffin-embedded sections of biopsies obtained from psoriatic skin ( n = 10), including non-lesional (NLS) ( i ), proximal-to-lesion (Pre-LS) ( ii ) and lesional (LS) zones of evolving plaques ( iii ), as well as from healthy donors ( n = 6) ( iv ) and AD skin ( n = 6) ( v ). Sections were counterstained with Mayer’s H&E. Bars, 100 μm. ( B ) Graphs show the mean of four-stage score values for IGFBP2, p16 and p-p21 epidermal expression, or the mean of the number of p21-positive cells ± SD. Two different sections were analysed for each staining, and the positivity was evaluated in five adjacent fields. ( C ) mRNA expression of IGFBP2, p16 and p21 was analysed by Real-time PCR on total RNA from healthy, NLS, LS biopsies ( n = 6) and normalized to GAPDH levels. The results are shown as individual values, mean and ± SD of relative mRNA levels. In ( B , C ), * p ≤ 0.05, p ** ≤ 0.01, as calculated by Mann–Whitney U test.

Article Snippet: For the specific neutralization of IGFBP2 bioactivity in keratinocyte supernatants, the neutralizing goat polyclonal anti-human IGFBP2 Ab (AF674) or goat IgG control (AB-108-C) from R&D Systems was used.

Techniques: Staining, Expressing, Real-time Polymerase Chain Reaction, MANN-WHITNEY

Journal: Aging (Albany NY)

Article Title: Intracellular Insulin-like growth factor binding protein 2 (IGFBP2) contributes to the senescence of keratinocytes in psoriasis by stabilizing cytoplasmic p21

doi: 10.18632/aging.103045

Figure Lengend Snippet: IGFBP2 production is induced by IGF-1 and is functionally active in psoriatic keratinocytes. IGFBP2 release was analysed by ELISA in protein lysates and supernatants (sups) of untreated healthy and pso KC cultures ( A ) or in sups of healthy and pso KC stimulated or not with 10 ng/ml IGF-1 or IGF-2 for 24 hours. ( B ) Data are expressed as mean of ng/10 6 cells ± SD of three different experiments carried out on different strains ( n = 3). * p ≤ 0.05, as assessed by unpaired Student’s t test; ** p ≤ 0.01, as calculated by the Mann–Whitney U test, comparing IGFBP2 production between healthy and pso KC groups. ( C ) BrdU incorporation was evaluated in healthy KC grown on coverslips and treated with complete medium alone ( i ) or 10-fold concentrated sup. of healthy KC ( ii ) or of pso KC (Pso) ( iii ). In parallel, healthy KC were treated with IGF-1 (10 ng/ml) alone ( iv ), or in presence of 10-fold concentrated healthy KC sups ( v ), or pso KC sups ( vi ). In two experimental conditions, 1 μg/ml of neutralizing anti-IGFBP2 Ab (α-IGFBP2) was added ( vii , viii ). Images are relative to one of three independent experiments performed on three different healthy KC strains. ( D ) Graphs represent the number of BrdU-positive cells counted in high power fields and expressed as cells per area unit ± SD ( n [microscopic fields per slide] = 6); H, healthy KC sups; Pso, psoriatic KC sups. ( E ) CXCL8 mRNA levels were detected by Real-time PCR in healthy KC stimulated for 8 hours with IGF-1 (10 ng/ml) alone or in combination with M4, including IFN-γ (200 U/ml), TNF-α (50 ng/ml), IL-17A (50 ng/ml) and IL-22 (50 ng/ml), in presence or not of healthy (H) or psoriatic (Pso) supernatants. In two experimental settings, 1 μg/ml of neutralizing anti-IGFBP2 Ab (α-IGFBP2) was added. Data shown are means of relative mRNA expression (normalized to GAPDH) of three independent experiments. ( D , E ), * p ≤ 0.05, as calculated by unpaired Student’s t test.

Article Snippet: For the specific neutralization of IGFBP2 bioactivity in keratinocyte supernatants, the neutralizing goat polyclonal anti-human IGFBP2 Ab (AF674) or goat IgG control (AB-108-C) from R&D Systems was used.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, BrdU Incorporation Assay, Real-time Polymerase Chain Reaction, Expressing

Journal: Aging (Albany NY)

Article Title: Intracellular Insulin-like growth factor binding protein 2 (IGFBP2) contributes to the senescence of keratinocytes in psoriasis by stabilizing cytoplasmic p21

doi: 10.18632/aging.103045

Figure Lengend Snippet: IGFBP2 expression progressively increases during senescence changes in psoriatic KC. ( A ) Healthy and pso KC were serially subcultured until they undergo senescence, and analysed by immunocytochemistry for the expression of IGFBP2 at two distinct culture passages (P1 and P4) (upper panels). Healthy KC were not counterstained with haematoxylin in order to preserve faint IGFBP2-specific staining (upper panels). The activity of senescence-associated β-galactosidase (SA- β-gal) was detected by colorimetric staining (blue) in healthy and pso KC cultures at passage P1 and P4. Data are representative of three independent experiments performed on different healthy ( n = 3) and psoriatic ( n = 3) donors. Bars, 50 μm. The graph shows the means of the percentage of IGFBP2 positive cells or SA- β-gal positive cells ± SD, counted in two adjacent fields. ( B , C ) WB analysis was performed on total protein lysates from healthy and pso KC at different serial passages of culture, left untreated ( B ) or M4-treated ( C ), to detect IGFBP2, p16, p21 and p-p21 expression. β-actin was used as loading control. Graphs represent the means of the densitometric intensity (D.I.) ± SD of the bands obtained from three different WB experiments. * p ≤ 0.05, ** p ≤ 0.01, as calculated by unpaired Student’s t test, comparing healthy and pso KC groups, or M4-treated and untreated groups.

Article Snippet: For the specific neutralization of IGFBP2 bioactivity in keratinocyte supernatants, the neutralizing goat polyclonal anti-human IGFBP2 Ab (AF674) or goat IgG control (AB-108-C) from R&D Systems was used.

Techniques: Expressing, Immunocytochemistry, Staining, Activity Assay, Control

Journal: Aging (Albany NY)

Article Title: Intracellular Insulin-like growth factor binding protein 2 (IGFBP2) contributes to the senescence of keratinocytes in psoriasis by stabilizing cytoplasmic p21

doi: 10.18632/aging.103045

Figure Lengend Snippet: IGFBP2 co-localizes with p21 in the cytoplasm of senescent psoriatic keratinocytes and its expression is positively regulated by p16. ( A ) Immunofluorescence analysis was conducted on pso KC cultures ( n = 3) at passage 4 (P4) to evaluate IGFBP2, p16, p21 and p-p21 subcellular localization. Cells were immunostained with anti-IGFBP2 Ab, followed by Cy3-conjugated secondary Ab (orange, panel i) or by Alexa Fluor 488-conjugated secondary Ab (green, panel v, ix), or, alternatively with p16 followed by Alexa Fluor 488 secondary Ab (green, panel ii), and p21 or p-p21 primary antibodies followed by Alexa Fluor 555 secondary Ab (orange, panels vi, x). Nuclei were counterstained with DAPI (blue) and the merging of three patterns was shown within the same field (merge, panels iv, viii, xii). Bars, 100 μM ( B ) IGFBP2, p16, p21 and p-p21 expression was analysed by WB on nuclear and cytosolic protein fractions obtained from two different pso KC strains (pso KC s1; pso KC s2), left untreated or treated with M4 for 18 hours. The quality of nuclear and cytosolic fractions was assessed by detection of lamin A/C and α-tubulin, respectively. ( C ) WB analysis was performed on lysates from primary human KC transduced with empty vector (mock) or p16 sense (p16S) vector and analysed for p16 and IGFBP2 expression. ( D ) Similarly, p16 and IGFBP2 expression was evaluated by WB on protein lysates obtained from primary KC cultures transduced with empty (mock) or antisense (p16AS). In ( C , D ), graphs show the means of the densitometric intensity (D.I.) of the bands ± SD, obtained from three independent experiments. * p ≤ 0.05, as calculated by Mann–Whitney U test.

Article Snippet: For the specific neutralization of IGFBP2 bioactivity in keratinocyte supernatants, the neutralizing goat polyclonal anti-human IGFBP2 Ab (AF674) or goat IgG control (AB-108-C) from R&D Systems was used.

Techniques: Expressing, Immunofluorescence, Transduction, Plasmid Preparation, MANN-WHITNEY

Journal: Aging (Albany NY)

Article Title: Intracellular Insulin-like growth factor binding protein 2 (IGFBP2) contributes to the senescence of keratinocytes in psoriasis by stabilizing cytoplasmic p21

doi: 10.18632/aging.103045

Figure Lengend Snippet: IGFBP2 interacts with p21 and protects it from ubiquitin-mediated proteasome degradation. ( A ) IGFBP2, p53, p21, p-p21, p27 and p16 protein expression was detected by WB analysis in pso KC cultured at passage 4 (P4, senescent cells) and silenced (si-IGFBP2) or not for IGFBP2 (si-NC) for different time points (right panels). Similarly, cyclin D, p-Rb and PCNA protein expression was detected in pso KC cultured at passage 1 (P1, pre-senescent cells) and silenced or not for IGFBP2 (left panels). In A and B, graphs show the mean ± SD of densitometric intensity (D.I.) of three independent experiments. * p ≤ 0.05, ** p ≤ 0.01 as calculated by paired Student’s t test comparing si-IGFBP2 with si-NC. ( B ) Co-immunoprecipitation experiments were performed on protein lysates obtained from pso KC left untreated or treated by M4 for 6 hours and then immunoprecipitated with antibodies against IGFBP2 or goat IgG as negative control (IP: IGFBP2, left panel), and with p21 or mouse IgG (IP: p21, right panel). The immunoprecipitates were probed with anti-IGFBP2, -p21 or –p16 antibodies, as shown in left and right panels. WB analysis was also performed on cell lysates (Input) to detect IGFBP2 and p21 levels. Figures are representative of three independent experiments. ( C ) Protein extracts of three distinct pso KC strains, transfected with si-IGFBP2 or si-NC for 24 h and then treated or not with 20 μM of the proteasome inhibitor MG132 for 6 h, were subjected to WB for the detection of IGFBP2, p21 and p16 expression. WB panels are representative of three independent experiments and D.I. indicates values of densitometric intensity. ( D ) Endogenous p21 was immunoprecipitated by protein lysates obtained from pso KC cultures silenced or not for IGFBP2 for 24 h and treated with 20 μM of MG132 for 6 hours. WB analysis was performed for the detection of p21 ubiquitination by using anti-ubiquitin antibody. WB was also performed on cell lysates (Input) to detect IGFBP2 and p21 levels, as well as β-actin as loading control.

Article Snippet: For the specific neutralization of IGFBP2 bioactivity in keratinocyte supernatants, the neutralizing goat polyclonal anti-human IGFBP2 Ab (AF674) or goat IgG control (AB-108-C) from R&D Systems was used.

Techniques: Ubiquitin Proteomics, Expressing, Cell Culture, Immunoprecipitation, Negative Control, Transfection, Control

Journal: Aging (Albany NY)

Article Title: Intracellular Insulin-like growth factor binding protein 2 (IGFBP2) contributes to the senescence of keratinocytes in psoriasis by stabilizing cytoplasmic p21

doi: 10.18632/aging.103045

Figure Lengend Snippet: IGFBP2 suppression leads to enhanced apoptosis of senescent psoriatic keratinocytes. ( A ) Pso KC and healthy KC cultures at passage P4 were silenced or not for IGFBP2 or p21 for 48 hours, and treated or not with M4 for 24 hours. Apoptosis was examined by measuring Annexin V/PI fluorescent staining through FACS analysis. Graphs show the mean ± SD of the percentage of AnnV/PI double-positive cells of three independent experiments. * p ≤ 0.05, as calculated by paired Student’s t test comparing si-IGFBP2 or si-p21 with si-NC, or untreated with M4. ( B ) Protein lysates of untreated healthy KC and pso KC, silenced or not for IGFBP2 or p21, were analysed by WB to confirm IGFBP2 or p21 silencing. Graphs show the mean ± SD of densitometric intensity (D.I.) of three independent experiments. * p ≤ 0.05, ** p ≤ 0.01 as calculated by paired Student’s t test comparing si-IGFBP2 or si-p21 with si-NC. ( C ) Protein extracts of pso KC at passage 4, silenced or not for IGFBP2, or p21 for 48 hours, and stimulated or not with M4 for 24 hours, were subjected to WB for the detection of IGFBP2, p21, p-p21, procaspase 3, p-JNK and JNK protein expression. WB panels are representative of three independent experiments and graphs show the mean ± SD of densitometric intensity (D.I.) of three independent experiments. * p ≤ 0.05, ** p ≤ 0.01 as calculated by paired Student’s t test comparing si-IGFBP2 or si-p21 with si-NC.

Article Snippet: For the specific neutralization of IGFBP2 bioactivity in keratinocyte supernatants, the neutralizing goat polyclonal anti-human IGFBP2 Ab (AF674) or goat IgG control (AB-108-C) from R&D Systems was used.

Techniques: Staining, Expressing

Journal: Aging (Albany NY)

Article Title: Intracellular Insulin-like growth factor binding protein 2 (IGFBP2) contributes to the senescence of keratinocytes in psoriasis by stabilizing cytoplasmic p21

doi: 10.18632/aging.103045

Figure Lengend Snippet: Proposed integrated model of the intracellular and extracellular functions of IGFBP2 in psoriasis keratinocytes. In epidermal keratinocytes of psoriasis patients characterized by a pre-senescent state, IGFBP2, p21 and p16 co-localize in the nuclear compartment. Here, p21 and p16 contribute to the cell cycle arrest in phase G1, by inhibiting Cdk1/2 and Cdk4/6, respectively. The nuclear role of IGFBP2 in pre-senescent KC remains to be investigated. In senescent psoriasis KC, IGFBP2, p16 and, of note, p21 accumulate in the cytoplasm. Here, p21 is hyper-phosphorylated and inhibits apoptotic processes. In the cytoplasm, IGFBP2 physically interacts with p21 and protects it from proteasomal degradation, thus sustaining its levels, and, indirectly, contributing to the apoptosis resistance typical of affected KC. In parallel, IGFBP2 released extracellularly blocks the pro-proliferative action of IGFs by binding to it and impeding its recruitment to IGF receptor.

Article Snippet: For the specific neutralization of IGFBP2 bioactivity in keratinocyte supernatants, the neutralizing goat polyclonal anti-human IGFBP2 Ab (AF674) or goat IgG control (AB-108-C) from R&D Systems was used.

Techniques: Binding Assay