Journal: Nature Communications

Article Title: FRET imaging of glycoRNA on small extracellular vesicles enabling sensitive cancer diagnostics

doi: 10.1038/s41467-025-58490-2

Figure Lengend Snippet: a Schematic diagram of the in situ FRET imaging of sEV glycoRNA binding to Siglec proteins. Predicted binding locations of the ISHPs and Siglec-Fc proteins (e.g., Siglec-10) are highlighted. b The AlphaFold predicted structures of Siglec-10, Siglec-11, and P-selectin. pLDDT corresponds to the model’s prediction of its score on the local Distance Difference Test. AlphaFold produces a per-residue model confidence score (pLDDT) between 0 and 100. Some regions below 50 pLDDT may be unstructured in isolation. AlphaFold Protein Structure Database, https://alphafold.ebi.ac.uk/ . c FCM analysis of bead-bound sEVs pre-treated with the indicated enzymes or inhibitors, followed by staining with the Cy3-conjugated Siglec-10. The gated region indicates that the population shifted toward high Siglec-10 binding. d Quantitative analysis of the positivity rate (Cy3 channel) depicted in ( c ). e Validation of the specific binding of glycoRNAs to Siglec-10. sEVs were pre-treated with RNase A and analyzed by FRET imaging (top). Scale bars: 10 μm (raw); 2 μm (enlarged). Quantification of average cdFRET intensity (bottom). f Representative cdFRET images of glycoRNA captured by using Cy3-conjugated Siglec-11 and Cy3-conjugated P-selectin for FRET assay. Scale bars: 10 μm (raw); 2 μm (enlarged). The red numbers represent the average intensities of the cdFRET signals. Data in ( d and e ) are shown as mean ± s.d. Statistical significance in ( d and e ) was determined by a two-tailed Student’s t test. P- values are indicated in the charts. All data are representative of three independent experiments.

Article Snippet: Specifically, 2 μl of Cy3 reagent (1 mg ml −1 ) was mixed with 300 μl of 10 μg human Siglec-10 recombinant protein (6 × His tag C-Terminus, > 95%, Sangon Biotech Co., Ltd., D145080), 300 μl of 10 μg human Siglec-11 recombinant protein (E84A, K145Q, CHO, hFc; MCE; HY-P79127), or 300 μl of 10 μg P-selectin (HEK293, His; MCE; HY-P70803), and incubated for 30 min at 37 °C in the dark.

Techniques: In Situ, Imaging, Binding Assay, Residue, Isolation, Staining, Biomarker Discovery, Two Tailed Test

Journal: Molecular medicine reports

Article Title: Effects of the eukaryotic initiation factor 6 gene on expression levels of inflammatory mediators in M2 macrophages during scar repair.

doi: 10.3892/mmr.2016.5294

Figure Lengend Snippet: Figure 4. mRNA expression levels of TIMP‑2, MMP‑2 and VEGF in M2 macrophages. mRNA levels were determined by reverse transcription‑quan titative polymerase chain reaction analysis, and protein expression levels were determined by ELISA. Data are presented as the mean ± standard deviation. **P<0.05, compared with eIF6+/+ M2 macrophages. eIF6, eukaryotic initia tion factor 6; TIMP, tissue inhibitor of metalloproteinase‑2; MMP‑2, matrix metalloproteinase‑2; VEGF, vascular endothelial growth factor.

Article Snippet: The supernatant obtained from the culture medium of macrophages following stimulation with IL-4 for 24 h was collected, and the expression levels of VEGF (cat. no. EK0506), MMP-2 (cat. no. EK0511) and TIMP-2 (cat. no. EK0322) were detected using ELISA kits (Boster Biological Technology Co., Ltd., Wuhan, China), in strict accordance with the manufacturer's protocol.

Techniques: Expressing, Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: International Journal of Biological Sciences

Article Title: Lung Cancer Cell-intrinsic Asparagine Synthetase Potentiates Anti-Tumor Immunity via Modulating Immunogenicity and Facilitating Immune Remodeling in Metastatic Tumor-draining Lymph Nodes

doi: 10.7150/ijbs.114791

Figure Lengend Snippet: ASNS shapes the immune landscapes in metastatic TdLN and primary tumor site. Figure A-C . LN metastasis model was conducted on C57BL/6 mice with LLC-ASNS WT (n=6), ASNS C2A overexpression cells(n=6) and control group(n=6), primary tumor and popliteal lymph nodes were isolated at the end of the experiment, and primary tumor volume(B) and TdLN volume/tumor volume(C) was measured and analyzed. 5D-F . (D) Representative FACS profiles of CD8+T cells are shown. The percentage(E) and number(F) of CD8+ subset in TIL cells isolated from primary tumor is shown. 5G-I . (G) Representative FACS profiles of the co-expression pattern of CD44 and CD62L, or the co-expression pattern of TCF-1and TOX in CD8+ T cells are shown. The number(H) and percentage(I) of CD44+, CD44+CD62L+, CD44+CD62L-, Tsl and TTSM subset in CD8+T cells isolated from TdLN is shown. 5J-L . (J) Representative FACS profiles of the co-expression pattern of CD44 and CD62L, or the co-expression pattern of TCF-1and TOX in CD8+ T cells are shown. The number(K) and percentage(L) of CD44+, CD44+CD62L+, CD44+CD62L-, Tsl and TTSM subset in CD8+T cells isolated from primary tumor is shown.

Article Snippet: For mIHC, the mouse tissue slides were sequentially labeled with monoclonal antibodies against CD44 (Bosterbio, A00052), CD62L (Bosterbio, PB9389), CD8 (Bosterbio, A02236-1), Ovabumin (Abcam, ab181688), and TCF-1 (Cell Signaling Technology, 2203T).

Techniques: Over Expression, Control, Isolation, Expressing

Journal: International Journal of Biological Sciences

Article Title: Lung Cancer Cell-intrinsic Asparagine Synthetase Potentiates Anti-Tumor Immunity via Modulating Immunogenicity and Facilitating Immune Remodeling in Metastatic Tumor-draining Lymph Nodes

doi: 10.7150/ijbs.114791

Figure Lengend Snippet: ASNS-high-expression metastases generated lymphocyte niches enriched with activated T cells, memory T cells, Tsl and TTSM. Figure A-C . Representative immunofluorescence staining images of metastatic TdLNs from LN metastasis model. 6D . The number of CD8+ T cells in the metastasis locations within TdLNs (ASNS WT , n=6, ASNS C2A , n=5, and EV, n=4). 6E-F . The number(E) and percentage(F) of CD44+CD8+T cells among all CD8 T cells in the metastasis locations within TdLNs (ASNS WT , n=6, ASNS C2A , n=5, and EV, n=4). 6G-H . The number(G) and percentage(H) of Tsl cells among all CD8 T cells in the metastasis locations within TdLNs (ASNS WT , n=6, ASNS C2A , n=5, and EV, n=4). 6I-J . The number(I) and percentage(J) of TTSM cells among all CD8 T cells in the metastasis locations within TdLNs (ASNS WT , n=5, ASNS C2A , n=4, and EV, n=3). 6K . Quantitative estimates of the distance from ova+ to CD8+CD44+CD62L+TCF+(TTSM) (ASNS WT , n=6, ASNS C2A , n=5, and EV, n=4). 6L-M . Representative immunofluorescence staining images of metastatic TdLNs from NSCLC patients. 6N-O . The number(C) and percentage(D) of CD8+ T cells in the metastasis locations within TdLNs(ASNS high group, n=7, and ASNS low group, n=6). 6P-Q . The number (E) and percentage(F) of CD45RO+CD8+ T cells in CD8+T cells in the metastasis locations within TdLNs(ASNS high group, n=7, and ASNS low group, n=6). 6R-S . The number (G) and percentage(H) of Tsl cells in CD8+T cells in the metastasis locations within TdLNs(ASNS high group, n=7, and ASNS low group, n=6). 6T-U . The number (I) and percentage(J) of TTSM cells in CD8+T cells in the metastasis locations within TdLNs(ASNS high group, n=7, and ASNS low group, n=6).

Article Snippet: For mIHC, the mouse tissue slides were sequentially labeled with monoclonal antibodies against CD44 (Bosterbio, A00052), CD62L (Bosterbio, PB9389), CD8 (Bosterbio, A02236-1), Ovabumin (Abcam, ab181688), and TCF-1 (Cell Signaling Technology, 2203T).

Techniques: Expressing, Generated, Immunofluorescence, Staining

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Vascular Protective Effects of Malus toringoides (Rehd.) Hughes Extracts and their Mechanism in Diabetic Rats and HUVECs

doi: 10.1155/2022/4348435

Figure Lengend Snippet: Effects of EMT on adhesion molecule expression in rat aortic histological sections and HUVECs. (a) Immunohistochemical analysis of adhesion molecule expression in aortic sections from a diabetic rat model established for 6 weeks and administered treatment for 8 weeks. (b–e) HUVECs were pretreated for 12 h with the indicated concentrations of EMT and stimulated for 48 h with high glucose (25 mM). ELISA and Western blot analyses of ICAM-1, VCAM-1, and E-selectin were performed as described in the materials and methods section. The values are mean ± SD of 3 independent experiments with triplicate samples. # P < 0.05 or ## P < 0.01 compared to the group treated with high glucose.

Article Snippet: ELISA kits for C-reactive protein (CRP, EK0978) tumor necrosis factor alpha (TNF- α , EK0526), interleukin 1 β (IL-1 β , EK0393), IL-6 (EK0412), VCAM-1 (EK0537), ICAM-1 (EK0370), E-selectin (EK0501) and CX3CL1 (EK0356) were obtained from BOSTER Biological Technology (Wuhan, China).

Techniques: Expressing, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Vascular Protective Effects of Malus toringoides (Rehd.) Hughes Extracts and their Mechanism in Diabetic Rats and HUVECs

doi: 10.1155/2022/4348435

Figure Lengend Snippet: Effects of the EMT components phlorizin, isoquercitrin, and quercitrin on the expression of CAMs in high glucose-induced HUVECs. HUVECs were pretreated for 12 h with the indicated concentrations of phlorizin, isoquercitrin, and quercitrin and then stimulated for 48 h with high glucose (25 mM). ELISA analyses of ICAM-1, VCAM-1, and E-selectin were performed as described in the materials and methods section. The values are mean ± SD of 3 independent experiments with triplicate samples. # P < 0.05 or ## P < 0.01 compared to the control group. ∗ P < 0.05 or ∗∗ P < 0.01 compared to the group treated with high glucose.

Article Snippet: ELISA kits for C-reactive protein (CRP, EK0978) tumor necrosis factor alpha (TNF- α , EK0526), interleukin 1 β (IL-1 β , EK0393), IL-6 (EK0412), VCAM-1 (EK0537), ICAM-1 (EK0370), E-selectin (EK0501) and CX3CL1 (EK0356) were obtained from BOSTER Biological Technology (Wuhan, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay