Journal: The Journal of Biological Chemistry

Article Title: Apolipoprotein E (APOE) regulates the transport of monosialotetrahexosylganglioside (GM1)

doi: 10.1016/j.jbc.2025.110778

Figure Lengend Snippet: Western blot results of APOE receptors: LDLR, VLDLR, LRP1, and ApoER2 on differentiated PC-12 cells, U-87 MG, bEnd.3, and HEK-293 cells. A , the expression of LDLR on differentiated PC-12 cells, U-87 MG, bEnd.3, and HEK-293 cells (n = 4). B , the expression of VLDLR on differentiated PC-12 cells, U-87 MG, bEnd.3, and HEK-293 cells (n = 4). C , the expression of LRP1 on differentiated PC-12 cells, U-87 MG, bEnd.3, and HEK-293 cells (n = 3). D , the expression of ApoER2 on differentiated PC-12 cells, U-87 MG, bEnd.3, and HEK-293 cells (n = 3). E , schematic illustration of determining the changes of APOE secondary structures. F , the CD results of APOE 3 under the effect of different GM1 content on the lipid structures (n ≥ 3). G , the CD results of APOE 4 under the effect of different GM1 content on the lipid structures (n = 3). p value: ns (0.05 < p ≤ 1), ∗ (0.01 < p ≤ 0.05, ∗∗ (0.001 < p ≤ 0.01, ∗∗∗ (0.0001 < p ≤ 0.001, ∗∗∗∗ ( p ≤ 0.0001). ( E ) is created with BioRender.com . APOE, apolipoprotein E; ApoER2, APOE receptor 2; HEK, human embryonic kidney; LDLR, low-density lipoprotein receptor; LRP1, LRPR-related protein 1; PC, l -α-phosphatidylcholine; VLDLR, very low-density lipoprotein receptor.

Article Snippet: To determine the binding affinity between APOE-lipoprotein and its receptor LDLR (MedChemExpress), we first labeled APOE3 and APOE4 with RED-NHS 2nd generation, as described earlier.

Techniques: Western Blot, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Apolipoprotein E (APOE) regulates the transport of monosialotetrahexosylganglioside (GM1)

doi: 10.1016/j.jbc.2025.110778

Figure Lengend Snippet: The binding affinity of APOE-enriched lipoprotein and APOE receptor LDLR using MST. A and B , the binding affinity between APOE3-enriched lipoprotein with varying GM1 concentration to LDLR. (n = 3) ( C and D ) the binding affinity between APOE4-enriched lipoprotein varying GM1 concentration to LDLR (n = 3). p value: ns (0.05 < p ≤ 1), ∗ (0.01 < p ≤ 0.05, ∗∗ (0.001 < p ≤ 0.01, ∗∗∗ (0.0001 < p ≤ 0.001, ∗∗∗∗ ( p ≤ 0.0001). APOE, apolipoprotein E; LDLR, low-density lipoprotein receptor; MST, microscale thermophoresis.

Article Snippet: To determine the binding affinity between APOE-lipoprotein and its receptor LDLR (MedChemExpress), we first labeled APOE3 and APOE4 with RED-NHS 2nd generation, as described earlier.

Techniques: Binding Assay, Concentration Assay, Microscale Thermophoresis

Journal: International immunopharmacology

Article Title: MiR-625-5p is a potential therapeutic target in sepsis by regulating CXCL16/CXCR6 axis and endothelial barrier.

doi: 10.1016/j.intimp.2024.112508

Figure Lengend Snippet: Fig. 1. Predictive value of biomarkers and clinical variables on cumulative ICU survival in patients with sepsis or septic shock patients. Blood samples were collected from patients withsepsis or septic shock and healthy subjects (control) for qRT-PCR analysis of miR-625-5p. miR-625-5p was normalized with the level of h-SNORD44 (internal control) to determine the ratios, and the ratios of the control were arbitrarily set at 1. The relative levels of miR-625-5p (a) and CXCL16 (b) were detected by qRT-PCR. The levels of CXCL16 (c), SDC-1 (d), HS (e), and VE-cadherin (f) were detected by using ELISA. The red rulers in Figures a and d represent mean values with SDs, quantitative data was compared with one-way ANOVA between three groups. The blue rulers in Figures b, c, e, and f represent the medians with ranges, quantitative data was compared with Kruskal–Wallis analysis between three groups. ROC curve analysis of APACHE II score, SOFA score, miR-625-5p, CXCL16, SDC-1, PCT, and lactate at admission for the prediction of 28-day mortality (g). Kaplan–Meier survival estimates for all patients with sepsis or septic shock according to the respective level of miR-625-5p (miR-625-5p, cut-of: 25) (h). Correlations between biomarker levels and various clinical parameters (i–n). *P < 0.05, **P < 0.01.

Article Snippet: Serum levels of syndecan-1 (EK1339, BOSTER, China), heparan sulfate (356350, Usbiological, USA), and VE-cadherin (DCADV0, R&D Systems, USA), and CXCL16 (EK0741, BOSTER, China) levels in the serum and cell supernatant were determined with an enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions.

Techniques: Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: International immunopharmacology

Article Title: MiR-625-5p is a potential therapeutic target in sepsis by regulating CXCL16/CXCR6 axis and endothelial barrier.

doi: 10.1016/j.intimp.2024.112508

Figure Lengend Snippet: Fig. 3. miR-625-5p regulated CXCL16 transcription and expression. EA.hy926 cells were transfected with miR-625-5p mimic (100 nM) or negative control (miR- 625-5p mimic NC) for 48 h. qRT-PCR analysis of miRNA expression levels for miR-625-5p by transfection of miR-625-5p mimics (a). The relative level of CXCL16 was detected by qRT-PCR (b), CXCL16 supernatant levels were quantified by ELISA (c), whereas the expression levels in EA.hy926 cells were detected by Western blotting (d). Quantification of CXCL16 is illustrated in (e). *P < 0.05, **P < 0.01.

Article Snippet: Serum levels of syndecan-1 (EK1339, BOSTER, China), heparan sulfate (356350, Usbiological, USA), and VE-cadherin (DCADV0, R&D Systems, USA), and CXCL16 (EK0741, BOSTER, China) levels in the serum and cell supernatant were determined with an enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions.

Techniques: Expressing, Transfection, Negative Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: International immunopharmacology

Article Title: MiR-625-5p is a potential therapeutic target in sepsis by regulating CXCL16/CXCR6 axis and endothelial barrier.

doi: 10.1016/j.intimp.2024.112508

Figure Lengend Snippet: Fig. 4. Inhibitor of miR-625-5p attenuated LPS-induced EA.hy926 cell barrier injury. EA.hy926 cells were transfected with the miR-625-5p inhibitor (50 nM) or negative control (micrOFF inhibitor NC) for 48 h and then exposed to LPS (5 and 10 µg/mL) for 6 h. The relative level of CXCL16 was detected by qRT-PCR (a), the expression levels of CXCL16 in the EA.hy926 cells were detected by Western blotting (b), and CXCL16 supernatant levels were quantified by ELISA (c). Quantification of CXCL16 is illustrated in (d). The effect of LPS and miR-625-5p mimic on the permeability of EA.hy926 cell were assessed using FITC-dextran and TEER methods (e, f). *P < 0.05, **P < 0.01.

Article Snippet: Serum levels of syndecan-1 (EK1339, BOSTER, China), heparan sulfate (356350, Usbiological, USA), and VE-cadherin (DCADV0, R&D Systems, USA), and CXCL16 (EK0741, BOSTER, China) levels in the serum and cell supernatant were determined with an enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions.

Techniques: Transfection, Negative Control, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Permeability

Journal: International immunopharmacology

Article Title: MiR-625-5p is a potential therapeutic target in sepsis by regulating CXCL16/CXCR6 axis and endothelial barrier.

doi: 10.1016/j.intimp.2024.112508

Figure Lengend Snippet: Fig. 6. CXCL16 knockdown inhibited LPS-induced endothelial cell injury in vitro. EA.hy926 cells were transfected with CXCL16 siRNA (siCXCL16-1, 2, or 3) or negative control (siRNA NC) and CXCL16 expression was determined after 24 h. The relative level of CXCL16 were detected by qRT-PCR (a). EA.hy926 cells were divided into four groups: Group 1, control; Group 2, LPS; Group 3, LPS + siCXCL16; Group 4, siRNA NC. The cells in Group 3 were transfected with CXCL16 siRNA (siCXCL16), whereas cells in the other groups were transfected with negative control siRNA (siRNA NC). At 24 h post-transfection, the cells in Groups 2 and 3 were treated with 10 µg/mL LPS. Endothelial permeability was measured by using FITC- dextran and TEER 6 h after LPS treatment (b, c). The levels of HS, SDC-1, claudin5, occludin, and VE-cadherin were detected by Western blotting (d). Quantification of HS, SDC-1, claudin5, occludin, and VE-cadherin are shown in (e): *P < 0.05, vs. Control group; & P < 0.05, vs. LPS group. Immunofluorescence images of HS and SDC-1 in EA.hy926 cells (f, h; magnification, ×200; scale bar, 150 µm). Fluo rescence intensity analysis of g and i. *P < 0.05, **P < 0.01.

Article Snippet: Serum levels of syndecan-1 (EK1339, BOSTER, China), heparan sulfate (356350, Usbiological, USA), and VE-cadherin (DCADV0, R&D Systems, USA), and CXCL16 (EK0741, BOSTER, China) levels in the serum and cell supernatant were determined with an enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions.

Techniques: Knockdown, In Vitro, Transfection, Negative Control, Expressing, Quantitative RT-PCR, Control, Permeability, Western Blot, Immunofluorescence

Journal: International immunopharmacology

Article Title: MiR-625-5p is a potential therapeutic target in sepsis by regulating CXCL16/CXCR6 axis and endothelial barrier.

doi: 10.1016/j.intimp.2024.112508

Figure Lengend Snippet: Fig. 9. Mechanisms by which miR-625-5p disrupts lung endothelial barrier integrity. miR-625-5p level may be an effective biomarker for predicting 28-day mortality in patients with sepsis or septic shock. Furthermore, LPS-induced vascular endothelial hyper-permeability by regulating miR-625-5p/CXCL16/CXCR6 axis in sepsis. ① LPS increased miR‑625-5p expression in EA.hy926 cells; ② miR-625-5p positively regulated CXCL16 in EA.hy926 cells; ③ Treatment of EA.hy926 cells with LPS significantly increased CXCL16 release; ④ CXCL16 combines with CXCR6; ⑤ CXCR6 mediated the effects of CXCL16 on endothelial barrier dysfunction.

Article Snippet: Serum levels of syndecan-1 (EK1339, BOSTER, China), heparan sulfate (356350, Usbiological, USA), and VE-cadherin (DCADV0, R&D Systems, USA), and CXCL16 (EK0741, BOSTER, China) levels in the serum and cell supernatant were determined with an enzyme-linked immunosorbent assay (ELISA) kit following the manufacturer’s instructions.

Techniques: Biomarker Discovery, Permeability, Expressing

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 1 CXCL1 is highly expressed in tissues with UCC. Strong immunohistochemical staining of CXCL1 in tissues with stage I (A), II (B) and III (C), comparing non-cancerous tissues with weak staining (D), was detected in a uterine cervical tissue microarray. Arrows, CXCL1 expression in stroma

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Immunohistochemical staining, Staining, Microarray, Expressing

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 2 High expression of CXCL1 positively correlated with worse survival in patients with CESC. A The role of CXCL1 expression level on patient overall survival probability. B The role of CXCL1 expression level and race on patient overall survival probability. C The role of CXCL1 expression level and body weight on patient overall survival probability. D The role of CXCR2 expression level on patient overall survival probability. E The role of CXCR2 expression level and race on patient overall survival probability. F The role of CXCR2 expression level and body weight on patient overall survival probability

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Expressing

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 3 CXCL1 expression was positively related with cancer-associated chemokines expression in CESC. A The genes that positively correlated with CXCL1 expression. B Heatmap of expression associated assay of 25 most relevant genes. C Gene expression correlation assays between CXCL and CCL20 (C), CXCL8 (D) or CXCL3 (E)

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Expressing, Gene Expression

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 4 Exogenous CXCL1 facilitates the malignant behaviors of HeLa cells. A The role of different concentrations of exogenous CXCL1 on the proliferation of HeLa cells was tested by CCK-8 assay. B-C Cell migration ability was determined by transwell analysis after HeLa cells treatment with different concentrations of exogenous CXCL1. *P < 0.05, **P < 0.01

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: CCK-8 Assay, Migration

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 5 CXCL1 overexpression contributes to proliferation, migration and apoptosis of HeLa cells via a autocrine manner. A The protein expression of CXCL1 in the supernatant derived from HeLa cells overexpressing CXCL1 and their mock controls medium was assessed by ELISA assay. B The proliferation ability of HeLa cells overexpressing CXCL1 was significantly increased as compared to their mock controls. C-D The cell migration capacity was notably enhanced after CXCL1 overexpression in HeLa cells. *P < 0.05, **P < 0.01

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Over Expression, Migration, Expressing, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 6 Stromal cell-derived CXCL1 enhances oncogenic potential of HeLa cells via a paracrine manner. A CXCL1 protein level in PHM1-41 cells overexpressing CXCL1 and their mock controls-derived supernatant was determined using ELISA assay. B The growth ability of HeLa cells treatment with CM from PHM1-41 cells overexpressing CXCL1 and their mock controls was detected by CCK-8 assay. C-D The effect of various proportions of CM derived from PHM1-41 cells overexpressing CXCL1 and their mock controls on the HeLa cell migration was explored by transwell analysis. *P < 0.05, **P < 0.01

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Migration

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 7 ERK signal involves in CXCL1-mediated malignant phenotypes in HeLa cells. A Expression of ERK, p-ERK, Cyclin D1 and Bax at protein levels from HeLa cells overexpressing CXCL1 and their mock controls were assessed by western blotting. B The protein level of the p-ERK was quantitated and normalized to total ERK from western blotting. C Protein expression of ERK, Cyclin D1 and BAX were statistically analyzed from western blotting, and their bands were normalized to β-actin. D The proliferation inhibition rate of ERK inhibitor PD98059 on HeLa cells overexpressing CXCL1 and their mock controls was determined by CCK-8 assay. E–F The effect of ERK inhibitor PD98059 on migration inhibition rate of HeLa cells overexpressing CXCL1 and their mock controls was assessed by transwell analysis. *P < 0.05, **P < 0.01

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Expressing, Western Blot, Inhibition, CCK-8 Assay, Migration