Journal: Frontiers in Cardiovascular Medicine

Article Title: CTRP3 attenuates myocardial lipotoxicity via suppression of lipid accumulation, inflammation, apoptosis, and mitochondrial oxidative stress

doi: 10.3389/fcvm.2025.1575929

Figure Lengend Snippet: (A,B) Compared to the high-fat diet (HFD) group, supplementation with CTRP3 significantly improves cardiac function in mice, as evidenced by enhanced left ventricular ejection fraction. (C,D) Histological examination of myocardial tissue reveals substantial lipid droplet accumulation in the HFD group. However, CTRP3 supplementation reduces lipid droplet accumulation, improves myocardial tissue structure, and alleviates the degree of fibrosis. (E–M) Immunofluorescence results indicate that CTRP3 supplementation mitigates myocardial inflammation, apoptosis, and oxidative stress in mice. (N–Q) Serum ELISA assays demonstrate that levels of triglycerides (TG), total cholesterol (TCHO), inflammatory markers, apoptotic factors, and oxidative stress indicators are significantly improved in the CTRP3 group compared to the HFD group ( n ≥ 3, p < 0.05).

Article Snippet: The recombinant CTRP3 protein (rCTRP3, Catalog Number: CSB-EP883621HU) was commercially procured from Wuhan Cusabio Biotech Co., Ltd. (Hubei, China).

Techniques: Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cardiovascular Medicine

Article Title: CTRP3 attenuates myocardial lipotoxicity via suppression of lipid accumulation, inflammation, apoptosis, and mitochondrial oxidative stress

doi: 10.3389/fcvm.2025.1575929

Figure Lengend Snippet: (A–F) Lipid testing results demonstrate that palmitic acid (PA) induces lipid accumulation in myocardial cells, while CTRP3 supplementation significantly reduces lipid droplet accumulation. (G) CTRP3 intervention improves the expression of genes related to fatty acid uptake, fatty acid oxidation, and lipid efflux in myocardial cells ( n ≥ 3, p < 0.05). (H) Quantitative PCR (q-PCR) results indicate that CTRP3 mitigates inflammation, apoptosis, and oxidative stress in myocardial cells stimulated by PA ( n ≥ 3, p < 0.05).

Article Snippet: The recombinant CTRP3 protein (rCTRP3, Catalog Number: CSB-EP883621HU) was commercially procured from Wuhan Cusabio Biotech Co., Ltd. (Hubei, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Cardiovascular Medicine

Article Title: CTRP3 attenuates myocardial lipotoxicity via suppression of lipid accumulation, inflammation, apoptosis, and mitochondrial oxidative stress

doi: 10.3389/fcvm.2025.1575929

Figure Lengend Snippet: (A–E) RNA sequencing analysis of myocardial tissue from the control group, HFD group, and CTRP3 group indicates that CTRP3 likely exerts its effects within myocardial mitochondria, playing a role in the metabolic regulation of myocardial lipotoxicity.

Article Snippet: The recombinant CTRP3 protein (rCTRP3, Catalog Number: CSB-EP883621HU) was commercially procured from Wuhan Cusabio Biotech Co., Ltd. (Hubei, China).

Techniques: RNA Sequencing, Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: CTRP3 attenuates myocardial lipotoxicity via suppression of lipid accumulation, inflammation, apoptosis, and mitochondrial oxidative stress

doi: 10.3389/fcvm.2025.1575929

Figure Lengend Snippet: (A,B) JC-1 staining results demonstrate a decrease in mitochondrial membrane potential in myocardial cells of the PA group. (C,D) Mito-Tracker staining results show a reduction in mitochondrial number in the PA group. (E) PA intervention in myocardial cells leads to decreased ATP production, while CTRP3 intervention improves ATP generation. (F) Transmission electron microscopy reveals increased mitochondrial damage and autophagy in myocardial cells following PA intervention. CTRP3 can mitigate mitochondrial damage and autophagy in myocardial cells.

Article Snippet: The recombinant CTRP3 protein (rCTRP3, Catalog Number: CSB-EP883621HU) was commercially procured from Wuhan Cusabio Biotech Co., Ltd. (Hubei, China).

Techniques: Staining, Membrane, Transmission Assay, Electron Microscopy

Journal: Frontiers in immunology

Article Title: IL6/adiponectin/HMGB1 feedback loop mediates adipocyte and macrophage crosstalk and M2 polarization after myocardial infarction.

doi: 10.3389/fimmu.2024.1368516

Figure Lengend Snippet: FIGURE 3 Adipocyte-derived ADPN promoted macrophage M2a polarization and decreased inflammatory cytokine secretion via the ADPN/AdipoR2/HMGB1 axis. (A) Single-cell sequencing analysis demonstrated that ADPN was mainly expressed in adipocytes in the subepicardial layer while the two ADPN receptors were expressed in almost cells in heart. (B) The ADPN knockdown efficiency in adipocytes using qPCR and WB analysis. (C) The polarization-related mRNA levels in hypoxic macrophages were measured using qPCR analysis when treated with the conditional medium of ADPN knockdown or control adipocytes. CM, conditional medium. (D) Flow cytometry of CD206 and Edu staining to determine the M2 polarization in hypoxic macrophage when treated with the conditional medium of ADPN knockdown or control adipocytes. (E) WB analysis demonstrated HMGB1 and inflammatory response in normoxic and hypoxic macrophages when treated with ADPN or control. (F) After HMGB1 knockdown, ADPN effects on inflammatory response and autophagy were rescued in hypoxic macrophages. (G) WB analysis showed that HMGB1 and inflammatory response in hypoxic macrophages when treated with the conditional medium of adipocytes and binding with macrophage AdipoR1, AdipoR2, avb3, and avb5 using the antibodies. (H) HMGB1 knockdown or control RAW264.7 macrophages were exposed to hypoxia with ADPN treatment, then were fixed by high-pressure freezing and analyzed by TEM. Mitochondria–lysosome contacts (contacting) and megamitochondria engulfing lysosome (engulfing) were shown in different groups. L, lysosome, or lysosome-related organelle; M, mitochondrion. (I) The unfit materials ejection from macrophages using TEM. Arrow, migrasomes. Scale bar, 500 nm. (J) The IL6, HMGB1, and C1qTNF9 expressions secreted by hypoxic macrophages when treated with the conditional medium of ADPN knockdown or control adipocytes. ADPNi, ADPN knockdown in adipocytes. *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant.

Article Snippet: The concentrations of cytokines secreted by macrophages were measured using the mouse IL6 ELISA kit (ab222503), mouse HMGB1 ELISA kit (JL13702, Jianglaibio, China), and mouse C1qTNF9 ELISA kit (CSB-E16713h, Cusabio, China).

Techniques: Derivative Assay, Sequencing, Knockdown, Control, Flow Cytometry, Staining, Binding Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting CD93 on monocytes revitalizes antitumor immunity by enhancing the function and infiltration of CD8+ T cells

doi: 10.1136/jitc-2024-010148

Figure Lengend Snippet: CD93 + monocytes accumulate in the peritumor of human HCC and indicate worse patient survival. (A) Distribution of CD68 + CD93 + cells in HCC tissues was determined by IF staining. Red, CD93; Green, CD68; Light Gray, CD31; Blue, DAPI. Scale bar=50 µm, n=10. B-D, CD14 + monocytes were purified from non-tumor, peritumor, or tumor tissues of HCC. Levels of CD93 expression in/on these cells were analyzed by western blotting (B, n=3), flow cytometry (C, n=4), or Q-PCR (D, n=5). (E) CD93 expression by different cells within HCC tissues was measured using scRNA-seq data from an online database. (F) 99 HCC patients who underwent curative resection with follow-up data were divided into two groups according to the median value of the CD68 + or CD68 + CD93 + cell density in peritumoral tissues (CD68 + cells: low, ≤405.7 cells/mm 2 (n=49); high, >405.7 cells/mm 2 (n=50); CD68 + CD93 + cells: low, ≤22.1 cells/mm 2 (n=49); high, >22.1 cells/mm 2 (n=50)). The overall survival and tumor recurrence of these patients were analyzed via the Kaplan-Meier method and log-rank test. Results shown in A, C and D are expressed as mean±SEM. *p<0.05, **p<0.01. The following statistical analyses were performed: one-way ANOVA ( A, C and D ), or log-rank test ( F and G ). ANOVA, analysis of variance; HCC, hepatocellular carcinoma; IF, immunofluorescence.

Article Snippet: Medium, siNC-containing liposome, or si CD93 -containing liposome (C12-200, HY-145405, MCE; cholesterol, C8503, Sigma-Aldrich; DSPC, 850 365P, Avanti; DMG-PEG2000, 880 151P, Avanti) were prepared as previously described.

Techniques: Staining, Purification, Expressing, Western Blot, Flow Cytometry, Immunofluorescence

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting CD93 on monocytes revitalizes antitumor immunity by enhancing the function and infiltration of CD8+ T cells

doi: 10.1136/jitc-2024-010148

Figure Lengend Snippet: Glycolytic switch induces CD93 upregulation on monocytes via the Erk signaling pathway. CD14 + cells were purified from the peripheral blood of healthy donors. (A–C) Cells were left untreated (Med) or treated with HepG2 TSN. The kinetics of CD93 expression by these cells were determined via Q-PCR (A, n=3), flow cytometry (B, n=4) and western blotting (C, n=4). (D, E) CD14 + cells were left untreated (Med) or treated with HepG2 TSN for 12 hours (D, n=7) or 24 hours (E, n=3) in the presence or absence of 2DG (20 mM) or 3PO (20 µM). Their expression of CD93 was measured by Q-PCR and flow cytometry. (F, G) Cells were left untreated or treated with HepG2 TSN for 24 hours in the presence or absence of NF-κB inhibitor JSH-23 (10 µM), or STAT3 inhibitor AG490 (10 µM), or JNK inhibitor SP600125 (10 µM), or Erk inhibitor U0126 (25 µM), or p38 inhibitor SB203580 (25 µM). Their levels of CD93 expression were determined by western blotting (F, n=3) or flow cytometry (G, n=4). (H) Cells were left untreated or treated with HepG2 TSN for 30 min in the presence or absence of 2DG (20 mM). Their levels of p-Erk and Erk expression were determined by western blotting. n=3. Results shown in A, B, D, E, G are expressed as mean±SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. The following statistical analyses were performed: paired two-tailed Student’s t-test ( A, B ), one-way ANOVA ( D, E ), or two-way ANOVA ( G ). ANOVA, analysis of variance.

Article Snippet: Medium, siNC-containing liposome, or si CD93 -containing liposome (C12-200, HY-145405, MCE; cholesterol, C8503, Sigma-Aldrich; DSPC, 850 365P, Avanti; DMG-PEG2000, 880 151P, Avanti) were prepared as previously described.

Techniques: Purification, Expressing, Flow Cytometry, Western Blot, Two Tailed Test

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting CD93 on monocytes revitalizes antitumor immunity by enhancing the function and infiltration of CD8+ T cells

doi: 10.1136/jitc-2024-010148

Figure Lengend Snippet: CD93 mediates tumor-induced PD-L1 expression on monocytes. (A–E) CD14 + cells were purified from the peripheral blood of healthy donors. Cells were transfected with control siRNA (siNC) or si CD93 before being treated with medium (Med) or HepG2 TSN for 24 hours. Their levels of CD93 (A, n=4), IL-6, IL-1β, TNF-α (B, n=6), HLA-DR, CD86 (C, n=4) were determined by flow cytometry or ELISA. The expression levels of PD-L1 were determined by flow cytometry (D, n=6) and western blotting (E, n=4). (F) Frozen sections of HCC samples were stained with anti-human CD68 antibody (green), anti-human CD93 antibody (red), anti-human PD-L1 antibody (white), and DAPI (blue). The co-localization of cell signals in the peritumoral regions was analyzed by confocal microscopy. Scale bar=50 µm, n=3. G, CD14 + cells were purified from tumor tissues of nine patients with HCC. Correlations between the expression levels of CD93 and PD-L1 in these cells were analyzed via flow cytometry. Results shown in A–E are expressed as mean±SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. The following statistical analyses were performed: two-way ANOVA ( A–E ), or Pearson’s correlation and linear regression analysis ( G ). ANOVA, analysis of variance; HCC, hepatocellular carcinoma; IF, immunofluorescence.

Article Snippet: Medium, siNC-containing liposome, or si CD93 -containing liposome (C12-200, HY-145405, MCE; cholesterol, C8503, Sigma-Aldrich; DSPC, 850 365P, Avanti; DMG-PEG2000, 880 151P, Avanti) were prepared as previously described.

Techniques: Expressing, Purification, Transfection, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Staining, Confocal Microscopy, Immunofluorescence

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting CD93 on monocytes revitalizes antitumor immunity by enhancing the function and infiltration of CD8+ T cells

doi: 10.1136/jitc-2024-010148

Figure Lengend Snippet: CD93 increases levels of PD-L1 through the AKT-GSK3β signaling pathway. (A–E) CD14 + cells were purified from the peripheral blood of healthy donors. (A) Cells were transfected with siNC or si CD93 , and left untreated or treated with HepG2 TSN for 24 hours. Their levels of CD93, p-GSK3β, GSK3β, p-AKT, AKT, p-p38, p38, p-FAK, and FAK expression were determined by western blotting. n=4. (B–E) Cells were left untreated or treated with HepG2 TSN in the presence or absence of AKTi (5 µM) for 24 hours, or LiCl (20 mM) for 24 hours. Their levels of p-GSK3β, GSK3β, and p-AKT expression were determined by western blotting ( B, D ), and their PD-L1 expression was analyzed by western blotting (B, D, n=3) and flow cytometry (C, E, n=5). (F, G) CD14 + cells were purified from tumor tissues of nine patients with HCC. Correlations between levels of PD-L1 and p-AKT expression, or PD-L1 and p-GSK3β expression in these cells were analyzed by western blotting. Results shown in (C, E) are expressed as mean±SEM. *p<0.05, **p<0.01, ****p<0.0001. The following statistical analyses were performed: two-way ANOVA ( C, E ) or Pearson’s correlation and linear regression analysis ( F, G ). ANOVA, analysis of variance; HCC, hepatocellular carcinoma.

Article Snippet: Medium, siNC-containing liposome, or si CD93 -containing liposome (C12-200, HY-145405, MCE; cholesterol, C8503, Sigma-Aldrich; DSPC, 850 365P, Avanti; DMG-PEG2000, 880 151P, Avanti) were prepared as previously described.

Techniques: Purification, Transfection, Expressing, Western Blot, Flow Cytometry

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting CD93 on monocytes revitalizes antitumor immunity by enhancing the function and infiltration of CD8+ T cells

doi: 10.1136/jitc-2024-010148

Figure Lengend Snippet: CD93 enhances the production of versican by monocytes. (A) CD14 + cells were purified from the peripheral blood of healthy donors, transfected with siNC or si CD93 , and then treated with HepG2 TSN for 24 hours. RNA-seq was performed with these differently-treated cells. The first 16 most downregulated genes in the si CD93 group versus the siNC control were shown. n=2. (B) Online HCC scRNA-seq data analysis revealed 145 genes upregulated in CD93 + versus CD93 − macrophages. These genes were compared with the downregulated genes in si CD93 -treated versus siNC-treated monocytes from our RNA-seq data, and the two overlapped genes were shown. (C–E) CD14 + monocytes were purified from non-tumor, peritumor, or tumor tissues of HCC. Levels of VCAN expression in these cells were analyzed by Q-PCR (C, n=5) or western blotting (D, n=3). Correlations between the mRNA levels of VCAN and CD93 in these cells were analyzed by Q-PCR (E, n=10). (F) CD14 + cells were purified from the peripheral blood of healthy donors, then left untreated (Med) or treated with HepG2 TSN. The kinetics of VCAN expression by these cells were determined via Q-PCR. n=4. (G, H) CD14 + cells purified from the peripheral blood of healthy donors were transfected with control siRNA (siNC) or si CD93 before being treated with medium (Med) or HepG2 TSN for 24 hours. Their levels of VCAN expression were determined by Q-PCR (G, n=4) and western blotting (H, n=4). Results shown in (C, F, H) are expressed as mean±SEM. *p<0.05, **p<0.01, ****p<0.0001. The following statistical analyses were performed: one-way ANOVA ( C ), Pearson’s correlation and linear regression analysis ( E ), paired two-tailed Student’s t-test ( F ) or two-way ANOVA ( G, H ). ANOVA, analysis of variance; HCC, hepatocellular carcinoma.

Article Snippet: Medium, siNC-containing liposome, or si CD93 -containing liposome (C12-200, HY-145405, MCE; cholesterol, C8503, Sigma-Aldrich; DSPC, 850 365P, Avanti; DMG-PEG2000, 880 151P, Avanti) were prepared as previously described.

Techniques: Purification, Transfection, RNA Sequencing, Control, Expressing, Western Blot, Two Tailed Test

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting CD93 on monocytes revitalizes antitumor immunity by enhancing the function and infiltration of CD8+ T cells

doi: 10.1136/jitc-2024-010148

Figure Lengend Snippet: Monocyte-derived versican suppresses the migration of CD8 + T cells. (A) VCAN expression by different cells within HCC tissues was measured using scRNA-seq data from an online database. (B) Distribution of Versican + , CD68 + , α-SMA + , and CD8 + cells in HCC tumor tissues was analyzed by IF staining. Scale bar=50 µm, n=4. (C) Correlations between levels of VCAN expression and CD8 infiltration were analyzed using TCGA data. (D) Paraffin-embedded sections of human HCC tissues were stained with anti-human CD93 and anti-CD8 antibodies. The patients were then divided into two groups according to the median value of peritumoral numbers of CD93 + cells. The levels of CD8 + T cells infiltration in peritumoral tissues (right panel) and intratumoral tissues (left panel) were compared between the two groups (CD93 Low , n=6; CD93 High , n=7). (E, F) CD14 + monocytes and CD8 + T cells were purified from the peripheral blood of healthy donors. Membrane of the 24-well Boyden chamber was coated with collagen I (1 mg/mL) plus hyaluronan (50 µg/mL), in the presence or absence of versican (10 µg/mL) ( E ), or the CCM or TCM from siNC or si VCAN -treated monocytes ( F ). CD8 + T cells were added to the upper compartment of the chamber, and their transmembrane migration was measured after 24 hours of incubation. n=4. Results shown in (D–F) were expressed as mean±SEM. *p<0.05, ***p<0.001, ****p<0.0001. The following statistical analyses were performed: Pearson’s correlation and linear regression analysis ( C ), two-tailed Student’s t-test ( D, E ) or two-way ANOVA ( F ). ANOVA, analysis of variance; HCC, hepatocellular carcinoma; IF, immunofluorescence; TCGA, the Cancer Genome Atlas.

Article Snippet: Medium, siNC-containing liposome, or si CD93 -containing liposome (C12-200, HY-145405, MCE; cholesterol, C8503, Sigma-Aldrich; DSPC, 850 365P, Avanti; DMG-PEG2000, 880 151P, Avanti) were prepared as previously described.

Techniques: Derivative Assay, Migration, Expressing, Staining, Purification, Membrane, Incubation, Two Tailed Test, Immunofluorescence

Journal: Journal for Immunotherapy of Cancer

Article Title: Targeting CD93 on monocytes revitalizes antitumor immunity by enhancing the function and infiltration of CD8+ T cells

doi: 10.1136/jitc-2024-010148

Figure Lengend Snippet: Targeting CD93 + monocytes inhibits tumor progression and enhances tumor sensitivity to anti-PD-1 treatment in mice in vivo. (A) C57BL/6 mice with established orthotopic, Luminescence-positive, Hepa1-6 tumors were injected with siNC-containing liposome, or si CD93 -containing liposome at indicated times via tail vein. (B, C) Real-time tumor growth and tumor size were monitored. (n=4). (D) Levels of Versican + F4/80 + cells infiltration in peritumoral tissues were measured by IF staining (n=4). (E–G) The infiltration and levels of IFN-γ and TNF-α expression of/by CD8 + T cells were measured on day 16 via flow cytometry (n=4). (H) C57BL/6 mice with established orthotopic, Luminescence-positive, Hepa1-6 tumors were injected with siNC-containing liposome or si CD93 -containing liposome via tail vein in the presence or absence of IgG or anti-PD-1 antibodies at indicated times. (I, J) Real-time tumor growth and tumor size were monitored (n=4). (K–M) Tumor infiltration and levels of IFN-γ and TNF-α expression of/by CD8 + T cells were measured on day 16 via flow cytometry (n=4). Results shown in (B, D–G, I, K–M) were expressed as mean±SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. The following statistical analyses were performed: one-way ANOVA with Turkey’s correction ( D–G, K–M ), two-way ANOVA with Turkey’s correction ( B, I ). ANOVA, analysis of variance; IF, immunofluorescence.

Article Snippet: Medium, siNC-containing liposome, or si CD93 -containing liposome (C12-200, HY-145405, MCE; cholesterol, C8503, Sigma-Aldrich; DSPC, 850 365P, Avanti; DMG-PEG2000, 880 151P, Avanti) were prepared as previously described.

Techniques: In Vivo, Injection, Staining, Expressing, Flow Cytometry, Immunofluorescence

Journal: Nature Communications

Article Title: A comprehensive synthetic library of poly- N -acetyl glucosamines enabled vaccine against lethal challenges of Staphylococcus aureus

doi: 10.1038/s41467-024-47457-4

Figure Lengend Snippet: A Complement deposition tests were performed as described using pooled sera from rabbits ( n = 2 per group) immunized with mQβ–PNAG conjugates, the 5GlcNH 2 –TT conjugate, or from a sample of pooled normal human sera. Titers and 95% confidence intervals (CI) were determined by linear regression using log 10 values of the average of replicate serum dilutions to determine the X intercept and 95% CI when Y = 0.5 (OD 405 nm of ELISA plate reading). P values indicate the significance of the deviation of the slope of the titration curve from zero to identify sera with activity at P < 0.05. mQβ–PNAG10 and mQβ–PNAG26 conjugates were more potent than the mQβ–PNAG0 and 5GlcNH 2 –TT conjugate in inducing C1q deposition onto purified PNAG. Normal human serum had no significant C1q depositing activity in spite of having a binding titer to PNAG (see Fig. ) consistent with prior reports that naturally acquired human antibody to PNAG is not functional due to the inability to activate the complement pathway , . Titers were determined by simple linear regression. B Pooled sera from rabbits ( n = 2 per group) immunized with mQβ–PNAG conjugate led to significantly higher levels of opsonic killing activities against S. aureus cells. Three aliquots were prepared from each pooled serum and the individual values of the three aliquots were presented. Source data are provided as a file.

Article Snippet: Subsequently, affinity‐purified goat anti‐human complement C1q antibody (Cedarlane, Inc, CL7341AP, 100 μL, 1:1000 dilution in GVB) was added to each well and incubated for 1 h under ambient temperature.

Techniques: Enzyme-linked Immunosorbent Assay, Titration, Activity Assay, Purification, Binding Assay, Functional Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway

doi: 10.3389/fcell.2021.700854

Figure Lengend Snippet: Relative expression of CTRP1 in MCAO/R-treated rats. (A) Relative protein expression of CTRP1 in the cortex detected by Western blot, n = 4 per group. (B) Relative mRNA expression of CTRP1 at different times after MCAO/R in the cortex, n = 4 per group. (C) Relative mRNA expression and concentration of CTRP1 in the serum of MCAO/R rats detected by qRT-PCR and ELISA, n = 12 per group. **** p < 0.0001, *** p < 0.001, ** p < 0.001, * p < 0.05 vs. sham group.

Article Snippet: CTRP1 antibody (1:50) and GRP78 (1:80) were mixed with protein A/G beads (MCE) at 4°C overnight.

Techniques: Expressing, Western Blot, Concentration Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway

doi: 10.3389/fcell.2021.700854

Figure Lengend Snippet: The location and expression of CTRP1 in the brain analyzed by immunofluorescence ( n = 3). (A) The expression of CTRP1 in neuron in the cortex, NeuN was used to label neuron. CTRP1 expression was observed by fluorescence microscope and is shown by green fluorescence. NeuN expression is shown by red fluorescence. The nuclei were stained with DAPI and is shown by blue fluorescence. (B) The expression of CTRP1 in astroglia in the cortex. GFAP was used to label astroglia. CTRP1 expression is shown by red fluorescence. GFAP expression is shown by green fluorescence. (C) The expression of CTRP1 in microglia in the cortex. IBA1 was used to label microglia. CTRP1 expression is shown by green fluorescence. IBA1 expression is shown by red fluorescence. (D) The intensity of CTRP1 and NeuN in the cortex. The representative images were acquired under × 400 magnification, scale bars = 50 μm. **** p < 0.0001 vs. sham group.

Article Snippet: CTRP1 antibody (1:50) and GRP78 (1:80) were mixed with protein A/G beads (MCE) at 4°C overnight.

Techniques: Expressing, Immunofluorescence, Fluorescence, Microscopy, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway

doi: 10.3389/fcell.2021.700854

Figure Lengend Snippet: CTRP1 attenuated brain injury in MCAO/R-treated rats. (A) The infection efficiency of LV-CTRP1 in the cortex. n = 3 per group. (B,C) Representative images of TTC staining and quantitative analysis of infarct volume. n = 4 per group. (D) Zea-longe scores. n = 13 per group. (E) HE staining and relative positive cells in the cortex. n = 3 per group. The representative images were acquired under × 400 magnification, scale bars = 50 μm, × 200 magnification, scale bars = 100 μm. **** p < 0.0001, vs. sham group, ** p < 0.01 vs. sham + LV-NC group, #### p < 0.0001, ### p < 0.001 vs. MCAO/R + LV-NC group.

Article Snippet: CTRP1 antibody (1:50) and GRP78 (1:80) were mixed with protein A/G beads (MCE) at 4°C overnight.

Techniques: Infection, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway

doi: 10.3389/fcell.2021.700854

Figure Lengend Snippet: CTRP1 protected against cerebral ischemia reperfusion injury via alleviating neuron injury and apoptosis. (A) Neuron injury was analyzed by double labeling immunofluorescence staining. n = 3 per group. The representative images were acquired under × 400 magnification, scale bars = 50 μm. (B) Apoptosis in the cortex was analyzed by TUNEL, n = 3 per group. The representative images were acquired under × 200 magnification, scale bars = 100 μm. (C) Western blot analyzed the expression of CTRP1, BAX, Bcl-2, and CHOP in cortex. n = 4 per group. **** p < 0.0001, *** p < 0.01, ** p < 0.01, * p < 0.05 vs. sham group; #### p < 0.0001, ## p < 0.01, # p < 0.05 vs. MCAO/R + LV-NC group.

Article Snippet: CTRP1 antibody (1:50) and GRP78 (1:80) were mixed with protein A/G beads (MCE) at 4°C overnight.

Techniques: Labeling, Immunofluorescence, Staining, TUNEL Assay, Western Blot, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway

doi: 10.3389/fcell.2021.700854

Figure Lengend Snippet: CTRP1 inhibited ERS via PERK signal pathway in the cortex of MCAO/R-treated rats. (A) Western blot analyzed the expression of PERK, p-PERK, GRP78, ATF6, ATF4, IRE1α, p-IRE1α. n = 4 per group. (B) CTRP1 affected the interaction between PERK and GRP78 after CIRI. n = 3 per group. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05 vs. sham group, #### p < 0.0001, ### p < 0.001, ## p < 0.01, # p < 0.05 vs. MCAO/R + LV-NC group.

Article Snippet: CTRP1 antibody (1:50) and GRP78 (1:80) were mixed with protein A/G beads (MCE) at 4°C overnight.

Techniques: Western Blot, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway

doi: 10.3389/fcell.2021.700854

Figure Lengend Snippet: The protective effect of CTRP1 in CIRI was abolished by CCT020312. (A,B) TTC staining and infarct volume in the brain. n = 4 per group. (C) Zea-longe scores. n = 13 per group. (D) HE staining and relative positive cells in the cortex. n = 3 per group. The representative images were acquired under × 200 magnification, scale bars = 100 μm, × 400 magnification, scale bars = 50 μm. (E) Double labeling immunofluorescence staining in the cortex (× 400 magnification). n = 3 per group. The representative images were acquired under × 400 magnification, scale bars = 50 μm. **** p < 0.0001, ∗∗ p < 0.01, * p < 0.05 vs. sham group, #### p < 0.0001, ### p < 0.001, ## p < 0.01, # p < 0.05 vs. MCAO/R + LV-NC group; @@ p < 0.01, @ p < 0.05 vs. MCAO/R + LV-CTRP1 group.

Article Snippet: CTRP1 antibody (1:50) and GRP78 (1:80) were mixed with protein A/G beads (MCE) at 4°C overnight.

Techniques: Staining, Labeling, Immunofluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway

doi: 10.3389/fcell.2021.700854

Figure Lengend Snippet: The function of CTRP1 on apoptosis and PERK signal pathway was inhibited by CCT020312. (A) Apoptosis was analyzed by TUNEL, n = 3 per group. The representative images were acquired under × 200 magnification, scale bars = 100 μm. (B) Western blot analyzed the expression CTRP1, BAX, CHOP, GRP78, ATF4, p-PERK, PERK, p-eIF2α, and eIF2α in the cortex. n = 6 per group. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05 vs. sham group, #### p < 0.0001, ## p < 0.01, # p < 0.05 vs. MCAO/R + LV-NC group, @@ p < 0.01, @ p < 0.05 vs. MCAO/R + LV-CTRP1 group.

Article Snippet: CTRP1 antibody (1:50) and GRP78 (1:80) were mixed with protein A/G beads (MCE) at 4°C overnight.

Techniques: TUNEL Assay, Western Blot, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway

doi: 10.3389/fcell.2021.700854

Figure Lengend Snippet: The function of CTRP1 in apoptosis and PERK signal pathway in neurons after OGD/R. (A) The expression of CTRP1 in neurons was analyzed by Western blot. n = 4 per group. (B) qRT-PCR analyzed the mRNA expression of CTRP1 after being transfected with siRNA and overexpression lentivirus, respectively. n = 3 per group. (C) Cell viability of neurons analyzed by MTT, n = 4 per group. (D) Apoptosis was analyzed by TUNEL, n = 3 per group. The representative images were acquired under × 200 magnification, scale bars = 100 μm. (E) The expression of CTRP1, BAX, Bcl-2, Cleaved caspase3, and Caspase 12 was analyzed by Western blot. n = 4 per group. (F) The protein expression of CHOP, GRP78, ATF4, PERK, and p-PERK in neurons. n = 4 per group. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05 vs. control group, #### p < 0.0001, ### p < 0.001, ## p < 0.01, # p < 0.05 vs. OGD/R + si-NC group, @@@@ p < 0.0001, @@@ p < 0.001, @ p < 0.05 vs. OGD/R + LV-NC group.

Article Snippet: CTRP1 antibody (1:50) and GRP78 (1:80) were mixed with protein A/G beads (MCE) at 4°C overnight.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Transfection, Over Expression, TUNEL Assay, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway

doi: 10.3389/fcell.2021.700854

Figure Lengend Snippet: CCT020312 reversed the effect of CTRP1 on apoptosis and the PERK signal pathway after OGD/R. (A) MTT analyzed the cell viability in neurons. n = 4 per group. (B) TUNEL analyzed apoptosis in neurons. n = 3 per group. The representative images were acquired under × 200 magnification, scale bars = 100 μm. (C) The expression of CTRP1, BAX, Bcl-2, Cleaved caspase 3, Caspase 12 was analyzed by Western blot. n = 4 per group. (D) The relative expression of CHOP, GRP78, ATF4, PERK and p-PERK in neurons. n = 4 per group. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05 vs. control group, #### p < 0.0001, ### p < 0.001, ## p < 0.01, # p < 0.05 vs. OGD/R + LV-NC group, @@@@ p < 0.0001, @@ p < 0.01, @ p < 0.05 vs. OGD/R + LV-CTRP1 group.

Article Snippet: CTRP1 antibody (1:50) and GRP78 (1:80) were mixed with protein A/G beads (MCE) at 4°C overnight.

Techniques: TUNEL Assay, Expressing, Western Blot, Control

Journal: Nature Communications

Article Title: Medium from human iPSC-derived primitive macrophages promotes adult cardiomyocyte proliferation and cardiac regeneration

doi: 10.1038/s41467-025-58301-8

Figure Lengend Snippet: a Immunofluorescence staining of Ki67-positive human cardiomyocytes treated with FM, hiPM-cm, hiPM-cm components retained by or passing through 3-kDa filters. Human cardiomyocytes were labeled with cTnT (red) and Ki67 (green); nuclei were counterstained with Hoechst (blue). Scale bar, 10 μm. n = 3 biological repeats per group. b Experimental strategy for analyzing protein components from hiPMpre-cm or hiPM-cm collected in RPMI medium by LC/MS. Cartoon elements used in the schematic diagram were created using BioRender. c Venn diagram showing the overlap between upregulated proteins identified in hiPM-cm versus hiPMpre-cm by LC/MS and upregulated genes encoding secreted proteins in CCR2 - human cardiac tissue-resident macrophages versus CCR2 + human monocyte-derived macrophages in the heart. d Unsupervised clustering analysis of relative pro-proliferative potentials of pairwise protein combinations evaluated by the percentages of Ki67-positive human cardiomyocytes. e Immunofluorescence staining of Ki67-positive human cardiomyocytes treated with a five-protein combination including C1QB, NRP1, PLTP, FUCA1, and SERPING1 (5-P combo). Scale bar, 10 μm. n = 3 biological repeats per group. f Principal component analysis of RNA sequencing data from human cardiomyocytes treated with FM and hiPM-cm. n = 3 biological repeats per group. g Volcano plot illustrating DEGs between human cardiomyocytes treated with FM and hiPM-cm. h Representative KEGG terms of cell proliferation-related genes upregulated in cardiomyocytes incubated with hiPM-cm versus FM. i Immunoblotting analysis showing the phosphorylated and total protein levels of AKT, ERK, and STAT3 in human cardiomyocytes treated with FM and hiPM-cm. GAPDH was used as a loading control. n = 3 biological repeats per group for phosphorylated and total ERK; n = 6 biological repeats per group for the other proteins. j Quantification of Ki67-positive human cardiomyocytes treated with hiPM-cm and inhibitors for PI3K-AKT (A-674563), ERK (PD0325901), and JAK-STAT (Stattic) pathways in single or in combination. n = 3 biological repeats per group. Quantitative data are presented as the mean ± SEM. Groups were compared using a two-tailed unpaired Student’s t test ( i ) or one-way ANOVA followed by post hoc Tukey test ( a , e , and j ). Source data are provided as a Source Data file.

Article Snippet: The proteins screened included: Transferrin (Novoprotein, CJ41), NRP2 (Sino Biological, 10695-H08H), SERPING1 (Novoprotein, C539), C1QC (Abclonal, RP01865), GAS6 (Novoprotein, C01W), C1QB (MCE, HY-P75464), FUCA1 (Sino Biological, 13893-H08H), NRP1 (Novoprotein, C11K), and PLTP (Sino Biological, 11171-H08H).

Techniques: Immunofluorescence, Staining, Labeling, Liquid Chromatography with Mass Spectroscopy, Derivative Assay, RNA Sequencing, Incubation, Western Blot, Control, Two Tailed Test