Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: IGFBP2 function as a novel biomarker for active lupus nephritis

doi: 10.1007/s00109-022-02241-z

Figure Lengend Snippet: TMT detection of differential protein expression between the active LN and HC groups. A Statistical histograms of GO ontology biological processes, cellular components, and molecular functions. B Bar graph displaying the top 10 enriched GO biological process functions. C Network analysis diagram. D Scatter plot of DE proteins and corresponding transcripts analyzed by the proteome and transcriptome. The x -axis is the transcript log 2 (fold-change), and the y -axis is the protein log 2 (fold-change). R = 0.0244 was the Pearson correlation coefficient of the two sets of data. The transcript was differentially expressed according to whether it was expressed in different colors. E Heatmap of proteins that were differentially expressed at the proteome and transcriptome levels and were consistently up- or downregulated. F Real-time PCR validation tests of IGFBP2 gene expression were performed in peripheral blood mononuclear cells (PBMCs) of the control ( n = 10) and LN ( n = 24) groups. ELISA validation tests were performed to verify IGFBP2 expression in the plasma of the controls ( n = 18), inactive LN ( n = 18), and active LN patients ( n = 23). G Validation of IGFBP2 expression in the PBMCs of HC and LN patients by WB ( n = 3). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: Briefly, protein samples (20 μg) were resolved by 12% SDS–PAGE, electroblotted on nitrocellulose membranes (Bio-Rad), incubated overnight at 4 °C with the following antibodies: IGFBP2 (11065–3-AP, Proteintech), antibodies used for Western blotting, were bought from AiFang biological and listed as follows: p-RPS6KB1(AF14512), RPS6KB1(AF11049), p-4E-BP1(AF01102), 4E-BP1(AF03855), p-AKT(AF00453), AKT(AF01499), p-mTOR (AF00658), mTOR (AF02824).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Biomarker Discovery, Gene Expression, Control, Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: IGFBP2 function as a novel biomarker for active lupus nephritis

doi: 10.1007/s00109-022-02241-z

Figure Lengend Snippet: The level of plasma IGFBP2 expression in LNs and correlation analysis with clinical indicators. A Level of IGFBP2 expression in the plasma of RA ( n = 16), SS ( n = 16), active LN ( n = 17), and control ( n = 18) patients. B The level of urine IGFBP2 expression in LN patients ( n = 12) was higheer than that in HC ( n = 12) patients ( P < 0.01). C There was no significant difference between urinary IGFBP2 in patients with inactive ( n = 19) and active SLE ( n = 23). D There was no correlation between plasma and urinary IGFBP2 levels in LN patients ( n = 26). E Correlation analysis between plasma IGFBP2 levels and SLE activity-related indicators. F Correlation analysis between the plasma IGFBP2 and SLEDAI score at 3 months after treatment and change in SLEDAI before and after treatment. G The ROC curve revealed that IGFBP2 showed an AUC value of 0.992, with a sensitivity of 0.909 and specificity of 1.0 (95% CI 0.974–1.0, P < 0.001) at the cutoff of 512 ng/mL between inactive and active LN. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. ns , not statistically significant

Article Snippet: Briefly, protein samples (20 μg) were resolved by 12% SDS–PAGE, electroblotted on nitrocellulose membranes (Bio-Rad), incubated overnight at 4 °C with the following antibodies: IGFBP2 (11065–3-AP, Proteintech), antibodies used for Western blotting, were bought from AiFang biological and listed as follows: p-RPS6KB1(AF14512), RPS6KB1(AF11049), p-4E-BP1(AF01102), 4E-BP1(AF03855), p-AKT(AF00453), AKT(AF01499), p-mTOR (AF00658), mTOR (AF02824).

Techniques: Clinical Proteomics, Expressing, Control, Activity Assay

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: IGFBP2 function as a novel biomarker for active lupus nephritis

doi: 10.1007/s00109-022-02241-z

Figure Lengend Snippet: The level of IGFBP2 expression in the kidneys of LN. A IGFBP2 expression in normal kidney issues and LNs by IF staining ( n = 3). B IHC staining of IGFBP2 in HC ( n = 3), MN ( n = 3), and II–V type LN kidneys ( n = 3). Original magnification, × 400. The positive area is quantitatively displayed as a histogram on the right. * P < 0.05; ** P < 0.01

Article Snippet: Briefly, protein samples (20 μg) were resolved by 12% SDS–PAGE, electroblotted on nitrocellulose membranes (Bio-Rad), incubated overnight at 4 °C with the following antibodies: IGFBP2 (11065–3-AP, Proteintech), antibodies used for Western blotting, were bought from AiFang biological and listed as follows: p-RPS6KB1(AF14512), RPS6KB1(AF11049), p-4E-BP1(AF01102), 4E-BP1(AF03855), p-AKT(AF00453), AKT(AF01499), p-mTOR (AF00658), mTOR (AF02824).

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: IGFBP2 function as a novel biomarker for active lupus nephritis

doi: 10.1007/s00109-022-02241-z

Figure Lengend Snippet: The distribution of IGFBP2 in the organs of mice. A The level of IGFBP2 expression in the blood of MRL mice (16 weeks) ( n = 4) was higher than that of C57BL/6 mice ( n = 3). B ELISA detection of the IGFBP2 content in the various organs of C57BL/6 and MRL/lpr mice revealed that in most organs of MRL/lpr mice, the protein content of IGFBP2 was higher than that of the C57BL/6 mice. C WB showed the expression of IGFBP2 in the lymph nodes of MRL/lpr and C57BL/6 mice ( n = 3). D IHC showed that the level of IGFBP2 in the lymph nodes of MRL/lpr was significantly higher than that in the lymph nodes of C57BL/6 mice (intracellular tan deposition) ( n = 3). * P < 0.05

Article Snippet: Briefly, protein samples (20 μg) were resolved by 12% SDS–PAGE, electroblotted on nitrocellulose membranes (Bio-Rad), incubated overnight at 4 °C with the following antibodies: IGFBP2 (11065–3-AP, Proteintech), antibodies used for Western blotting, were bought from AiFang biological and listed as follows: p-RPS6KB1(AF14512), RPS6KB1(AF11049), p-4E-BP1(AF01102), 4E-BP1(AF03855), p-AKT(AF00453), AKT(AF01499), p-mTOR (AF00658), mTOR (AF02824).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: IGFBP2 function as a novel biomarker for active lupus nephritis

doi: 10.1007/s00109-022-02241-z

Figure Lengend Snippet: The IGFBP2 blocking effect in mice following treatment. A MRL/lpr mice were divided into the PBS group ( n = 4), ISO group ( n = 4), and anti-IGFBP2 group (n = 4). The ELISA results showed that compared with PBS and ISO groups, the level of plasma IGFBP2 in the anti-IGFBP2 group was decreased. B PCR and C WB showed the level of renal IGFBP2 protein expression in each of the three groups. * P < 0.05; ** P < 0.01; *** P < 0.001. ns , not statistically significant

Article Snippet: Briefly, protein samples (20 μg) were resolved by 12% SDS–PAGE, electroblotted on nitrocellulose membranes (Bio-Rad), incubated overnight at 4 °C with the following antibodies: IGFBP2 (11065–3-AP, Proteintech), antibodies used for Western blotting, were bought from AiFang biological and listed as follows: p-RPS6KB1(AF14512), RPS6KB1(AF11049), p-4E-BP1(AF01102), 4E-BP1(AF03855), p-AKT(AF00453), AKT(AF01499), p-mTOR (AF00658), mTOR (AF02824).

Techniques: Blocking Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Expressing

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: IGFBP2 function as a novel biomarker for active lupus nephritis

doi: 10.1007/s00109-022-02241-z

Figure Lengend Snippet: Improvement of LN after a IGFBP2 blockade. A MRL/lpr mice were divided into the PBS group ( n = 4), ISO group ( n = 4), and anti-IGFBP2 group ( n = 4). The skin lesions in the anti-IGFBP2 group were significantly improved. B Compared with the PBS and ISO groups, the lymph/body weight ratio was decreased in the anti-IGFBP2 group. C Trend in the reduction of urine protein following treatment. D HE, PAS, and Masson staining (original magnification, × 400), quantified histogram of glomerulus, tubulointerstitium and perivascular pathology. * P < 0.05; ** P < 0.01

Article Snippet: Briefly, protein samples (20 μg) were resolved by 12% SDS–PAGE, electroblotted on nitrocellulose membranes (Bio-Rad), incubated overnight at 4 °C with the following antibodies: IGFBP2 (11065–3-AP, Proteintech), antibodies used for Western blotting, were bought from AiFang biological and listed as follows: p-RPS6KB1(AF14512), RPS6KB1(AF11049), p-4E-BP1(AF01102), 4E-BP1(AF03855), p-AKT(AF00453), AKT(AF01499), p-mTOR (AF00658), mTOR (AF02824).

Techniques: Staining

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: IGFBP2 function as a novel biomarker for active lupus nephritis

doi: 10.1007/s00109-022-02241-z

Figure Lengend Snippet: IGFBP2 blockade affects T cells and AKT/mTOR pathway. A Effect on peripheral blood T cell immunity by flow cytometry in PBS group ( n = 4), ISO group ( n = 4), and anti-IGFBP2 group ( n = 4). B Protein expression and phosphorylation levels of AKT, mTOR, RPS6KB1, and 4E-BPI in anti-IGFBP2 group ( n = 4) and ISO group ( n = 3) were detected by Western blotting. The quantitative analysis of proteins was performed using ImageJ software. * P < 0.05; ** P < 0.01; ns , not statistically significant

Article Snippet: Briefly, protein samples (20 μg) were resolved by 12% SDS–PAGE, electroblotted on nitrocellulose membranes (Bio-Rad), incubated overnight at 4 °C with the following antibodies: IGFBP2 (11065–3-AP, Proteintech), antibodies used for Western blotting, were bought from AiFang biological and listed as follows: p-RPS6KB1(AF14512), RPS6KB1(AF11049), p-4E-BP1(AF01102), 4E-BP1(AF03855), p-AKT(AF00453), AKT(AF01499), p-mTOR (AF00658), mTOR (AF02824).

Techniques: Flow Cytometry, Expressing, Phospho-proteomics, Western Blot, Software

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Icaritin-curcumol activates CD8 + T cells through regulation of gut microbiota and the DNMT1/IGFBP2 axis to suppress the development of prostate cancer

doi: 10.1186/s13046-024-03063-2

Figure Lengend Snippet: ICA-CUR inhibits the DNMT1/IGFBP2 pathway and activates the cytotoxic effect of CD8 + T cell. A The levels of IGFBP2 in serum were tested via ELISA. B The levels of DNMT1 and IGFBP2 in tumor tissues were tested via IHC (Magnification: ×100, scale bar = 100 μm; Magnification: ×400, scale bar = 25 μm). C The levels of DNMT1 and IGFBP2 in tumor tissues were detected by WB. D The protein changes of PD-L1, EGFR, STAT3, p-EGFR, and p-STAT3 in tumor tissues were detected by WB. E FCM was used to test the infiltration of CD8 + T cells (positivity of CD3 + CD8 + IFN-γ and CD3 + CD8 + Ki67 cells). F The levels of perforin, granzyme A, and B in sorted CD8 + T cells from tumor tissues were detected by RT-PCR. G The levels of IFN-γ and IFN-α in serum were tested via ELISA. * P < 0.05 vs. PCa

Article Snippet: According to the kit’s instructions, ELISA was utilized to evaluate levels of IGFBP2 (CSB-E04589m, CUSABIO), interferon-γ (IFN-γ) (KE10001, Proteintech), interferon-α (IFN-α) (MFNAS0, R&D Systems), Perforin (Cbic-E13429m, CUSABIO), Granzyme A (CSB-E08717m, CUSABIO, China) and Granzyme B (CSB-E08720m, CUSABIO, China).

Techniques: Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Icaritin-curcumol activates CD8 + T cells through regulation of gut microbiota and the DNMT1/IGFBP2 axis to suppress the development of prostate cancer

doi: 10.1186/s13046-024-03063-2

Figure Lengend Snippet: FMT from donors in the ICA-CUR treatment inhibits the development of PCa and activates the cytotoxic effects of CD8 + T cells. A Tumor imaging, volume, and weight measurements. B IF staining was utilized to examine changes in Ki67 expression in tumors (Magnification: ×400, scale bar = 25 μm). C The levels of IGFBP2 in serum were tested via ELISA. D The levels of DNMT1 and IGFBP2 in tumor tissues were detected by WB. E WB was used to detected the protein changes of PD-L1, EGFR, STAT3, p-EGFR, and p-STAT3 in tumor tissues. F The infiltration of CD8 + T cells in mouse tumor tissues (positivity of CD3 + CD8 + IFN-γ and CD3 + CD8 + Ki67 cells) was detected by FCM. G The expression of perforin, granzyme A, and B in sorted CD8 + T cells from tumor tissues was detected by RT-PCR. H The levels of IFN-γ and IFN-α in serum were tested via ELISA. * P < 0.05 vs. FMT-PCa

Article Snippet: According to the kit’s instructions, ELISA was utilized to evaluate levels of IGFBP2 (CSB-E04589m, CUSABIO), interferon-γ (IFN-γ) (KE10001, Proteintech), interferon-α (IFN-α) (MFNAS0, R&D Systems), Perforin (Cbic-E13429m, CUSABIO), Granzyme A (CSB-E08717m, CUSABIO, China) and Granzyme B (CSB-E08720m, CUSABIO, China).

Techniques: Imaging, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Icaritin-curcumol activates CD8 + T cells through regulation of gut microbiota and the DNMT1/IGFBP2 axis to suppress the development of prostate cancer

doi: 10.1186/s13046-024-03063-2

Figure Lengend Snippet: ICA-CUR inhibits tumor development and activates cytotoxic effects of CD8 + T cells by suppressing the SCFAs-IGFBP2 axis. A The levels of DNMT1 and IGFBP2 in tumor tissues were detected by WB. B The levels of IGFBP2 in serum were tested via ELISA. C The levels of DNMT1 and IGFBP2 in tumor tissues were detected by WB. * P < 0.05 vs. ICA + CUR. D Tumor imaging, volume, and weight measurements. E IF staining to examine changes in Ki67 expression in tumors (Magnification: ×400, scale bar = 25 μm). F ELISA was used to detect the levels of IGFBP2 in serum. G The protein changes of PD-L1, EGFR, STAT3, p-EGFR, and p-STAT3 in tumor tissues were detected by WB. H The infiltration of CD8 + T cells in mouse tumor tissues (positivity of CD3 + CD8 + IFN-γ and CD3 + CD8 + Ki67 cells) was detected by FCM. I The levels of perforin, granzyme A, and B in sorted CD8 + T cells from tumor tissues were detected by RT-PCR. J ELISA was utilized to detect the levels of IFN-γ and IFN-α in serum. * P < 0.05 vs. ICA + CUR + IgG, # P < 0.05 vs. ICA + CUR + anti-IGFBP2

Article Snippet: According to the kit’s instructions, ELISA was utilized to evaluate levels of IGFBP2 (CSB-E04589m, CUSABIO), interferon-γ (IFN-γ) (KE10001, Proteintech), interferon-α (IFN-α) (MFNAS0, R&D Systems), Perforin (Cbic-E13429m, CUSABIO), Granzyme A (CSB-E08717m, CUSABIO, China) and Granzyme B (CSB-E08720m, CUSABIO, China).

Techniques: Enzyme-linked Immunosorbent Assay, Imaging, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Icaritin-curcumol activates CD8 + T cells through regulation of gut microbiota and the DNMT1/IGFBP2 axis to suppress the development of prostate cancer

doi: 10.1186/s13046-024-03063-2

Figure Lengend Snippet: ICA-CUR inhibits the development of PCa, the DNMT1/IGFBP2 pathway, and activates cytotoxic effects of CD8 + T cells in vitro. A CCK-8 was utilized to assess the proliferation ability of cells. B Transwell was applied to measure the migration and invasion ability of cells. C The level of IGFBP2 in cells was tested via ELISA. D The levels of DNMT1 and IGFBP2 in cells were detected by WB. E WB was utilized to monitor protein changes of PD-L1, EGFR, STAT3, p-EGFR, and p-STAT3. * P < 0.05 vs. Control. F FCM analysis of CD8 + IFN-γ cells. G The expression of perforin, granzyme A, and B in sorted CD8 + T cells was detected by WB. H IL-2, IFN-γ, and IFN-α levels in supernatant were measured through ELISA. I Perforin and granzyme B levels in the supernatant were tested via ELISA. & P < 0.05 vs. RM-1 + T cells

Article Snippet: According to the kit’s instructions, ELISA was utilized to evaluate levels of IGFBP2 (CSB-E04589m, CUSABIO), interferon-γ (IFN-γ) (KE10001, Proteintech), interferon-α (IFN-α) (MFNAS0, R&D Systems), Perforin (Cbic-E13429m, CUSABIO), Granzyme A (CSB-E08717m, CUSABIO, China) and Granzyme B (CSB-E08720m, CUSABIO, China).

Techniques: In Vitro, CCK-8 Assay, Migration, Enzyme-linked Immunosorbent Assay, Control, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Icaritin-curcumol activates CD8 + T cells through regulation of gut microbiota and the DNMT1/IGFBP2 axis to suppress the development of prostate cancer

doi: 10.1186/s13046-024-03063-2

Figure Lengend Snippet: ICA-CUR inhibits the development of PCa and activates the cytotoxic effects of CD8 + T cells through the inhibition of the DNMT1/IGFBP2 pathway. A Transfection efficiency detection by WB. B CCK-8 was utilized to assess the proliferation ability of cells. C Cell migration and invasion ability detection by Transwell assay. D The levels of DNMT1 and IGFBP2 in cells were detected by WB. E WB was utilized to monitor protein changes of PD-L1, EGFR, STAT3, p-EGFR, and p-STAT3. F FCM analysis of CD3 + CD8 + IFN-γ cells. G The levels of perforin, granzyme A, and B in sorted CD8 + T cells were detected by RT-qPCR. H The levels of IL-2, IFN-γ, and IFN-α in serum. I perforin, granzyme A and B levels in the supernatant were tested via ELISA. * P < 0.05 vs. Control, # P < 0.05 vs. ICR + CUR + oe-NC, & P < 0.05 vs. ICR + CUR + oe-DNMT1 + si-NC

Article Snippet: According to the kit’s instructions, ELISA was utilized to evaluate levels of IGFBP2 (CSB-E04589m, CUSABIO), interferon-γ (IFN-γ) (KE10001, Proteintech), interferon-α (IFN-α) (MFNAS0, R&D Systems), Perforin (Cbic-E13429m, CUSABIO), Granzyme A (CSB-E08717m, CUSABIO, China) and Granzyme B (CSB-E08720m, CUSABIO, China).

Techniques: Inhibition, Transfection, CCK-8 Assay, Migration, Transwell Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Icaritin-curcumol activates CD8 + T cells through regulation of gut microbiota and the DNMT1/IGFBP2 axis to suppress the development of prostate cancer

doi: 10.1186/s13046-024-03063-2

Figure Lengend Snippet: Inhibiting DNMT1 could suppress PCa development and activate the cytotoxic effects of CD8 + T cells by inhibiting the IGFBP2/EGFR/STAT3/PD-L1 pathway. A Transfection efficiency detection by WB. B CCK-8 was utilized to assess the proliferation ability of cells. C Cell migration and invasion ability detection by Transwell assay. D The levels of DNMT1 and IGFBP2 were detected by WB. E , F WB was utilized to monitor protein changes of PD-L1, EGFR, STAT3, p-EGFR, p-STAT3, DNMT1, and IGFBP2. G FCM analysis of CD3 + CD8 + IFN-γ cells. H The levels of perforin, granzyme A, and B were detected by RT-PCR. I The levels of IL-2, IFN-γ, and IFN-α in serum. J Perforin, granzyme A, and B levels in the supernatant were tested via ELISA. * P < 0.05 vs. si-NC, # P < 0.05 vs. si-DNMT1 + oe-NC

Article Snippet: According to the kit’s instructions, ELISA was utilized to evaluate levels of IGFBP2 (CSB-E04589m, CUSABIO), interferon-γ (IFN-γ) (KE10001, Proteintech), interferon-α (IFN-α) (MFNAS0, R&D Systems), Perforin (Cbic-E13429m, CUSABIO), Granzyme A (CSB-E08717m, CUSABIO, China) and Granzyme B (CSB-E08720m, CUSABIO, China).

Techniques: Transfection, CCK-8 Assay, Migration, Transwell Assay, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Therapeutic effect of modified zengye decoction on primary Sjogren’s syndrome and its effect on plasma exosomal proteins

doi: 10.3389/fphar.2022.930638

Figure Lengend Snippet: The plasma exosomalDEPs before and after MZD treatment.

Article Snippet: The exosomal protein ceruloplasmin (CP), cartilage oligomeric matrix protein (COMP), insulin-like growth factor-binding protein 2 (IGFBP2), ficolin-2 (FCN2), and lipopolysaccharide-binding protein (LBP) concentrations were determined using ELISA kits supplied by EIAab Science Inc. (Wuhan, China) according to the instructions of the manufacturer.

Techniques: