Journal: Advanced Science

Article Title: Spatial Transcriptomics Reveals Transcriptomic and Immune Microenvironment Reprogramming during Thyroid Carcinoma Dedifferentiation

doi: 10.1002/advs.202506925

Figure Lengend Snippet: PDCD4 regulates the secretion of M2‐polarizing proteins in an eIF4A‐dependent manner. A) Schematic diagram of PDCD4–eIF4A‐mediated regulation of protein expression patterns. B) Co‐IP analysis showing the changes in the eIF4F complex (eIF4A–eIF4E–eIF4G) upon PDCD4 knockout or overexpression in BHT101 cells. C) WB analysis confirmed the knockdown efficiency of eIF4A using siRNA in TPC‐sg‐PDCD4 and BHT101. D,E) FCM analysis showing that inhibition of eIF4A reduced M2 macrophage polarization induced by TPC‐sg‐PDCD4 (D) and BHT101 (E) cells in THP‐1 polarization assays. F) In vivo experiments showed that the eIF4A inhibitor suppressed subcutaneous tumor growth in mice. G,H) FCM revealed that eIF4A inhibition decreased M2 macrophage infiltration (G) and increased M1 macrophage infiltration (H) in tumors. I–L) Proteomic and pathway enrichment analyses of intracellular proteins (I, K) and secreted proteins in the supernatant (J, L) after PDCD4 knockout in TPC‐1 cells. M,N) Polysome profiling analysis demonstrated that PDCD4 knockout affected the translational efficiency of specific mRNAs in TPC‐1 cells. Co‐IP : Co‐immunoprecipitation ; eIF4A : eukaryotic Initiation Factor 4A ; eIF4E : eukaryotic Initiation Factor 4E ; eIF4G : eukaryotic Initiation Factor 4G ; FCM : Flow cytometry; PDCD4 : Programmed Cell Death 4; WB : Western Blot. Significance in D‐F was determined using one‐way ANOVA and the Bonferroni multiple comparison test. Significance in G, H was determined using Unpaired t‐test * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The eIF4A inhibitor CR‐1‐31‐B (MCE, Cat#HY‐136453) was administered to mice at a dose of 2 mg kg −1 via intraperitoneal injection every other day in mouse drug administration experiments.

Techniques: Expressing, Co-Immunoprecipitation Assay, Knock-Out, Over Expression, Knockdown, Inhibition, In Vivo, Immunoprecipitation, Flow Cytometry, Western Blot, Comparison

Journal: International Journal of Clinical Practice

Article Title: Pleural Mesothelial Cells-Induced Monocytes to the Pleural Cavity through the Effect of C3 Lytic Products in Tuberculous Pleural Effusion

doi: 10.1155/2024/5544085

Figure Lengend Snippet: Figure 2: Te concentrations of complement pyrolysis products were found higher in pleural efusion than in plasma from TPE patients. (a) Complement pyrolysis products, including C3a, C3b, C3d, C5a, and opsonin receptors (CR1 and CR3) were detected in human tuberculosis pleural biopsy samples by immunohistochemistry. (Original magnifcation, ×200) (n 4). (b) Higher levels of complement pyrolysis products were found in pleural efusion than in plasma in TPE patients (n 20). Te concentrations of complement pyrolysis products in pleural fuid and plasma from TPE patients were measured by ELISA (n 20).

Article Snippet: To investigate whether CR1, CR3, and CXCL12 contributed to monocyte migration, blocking experiments were performed by mixing monocytes with anti-CR1 mAb (5 μg/ml, BM2364, Origene), anti-CR3 mAb (10 μg/ml, 101213, Biolegend) or rabbit IgG irrelevant isotype control.

Techniques: Clinical Proteomics, Immunohistochemistry, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Clinical Practice

Article Title: Pleural Mesothelial Cells-Induced Monocytes to the Pleural Cavity through the Effect of C3 Lytic Products in Tuberculous Pleural Effusion

doi: 10.1155/2024/5544085

Figure Lengend Snippet: Figure 4: Monocyte migration was inhibited by antibodies that blocked CR1 and CR3. (a) Representative colocalization of C3b-CR1 and C3d-CR3 in PMCs and monocytes from TPE. Te cells from TPE were immunostained with C3b/C3d (red) and CR1/CR3 (green), and the nuclei were stained with DAPI (blue) (n 4). (b) To evaluate the efect of C3b-CR1 and C3d-CR3 axes on monocytes, monocytes were seeded into the top chamber with or without anti-CR1 antibody or anti-CR3 antibody, and the supernatants from cultured PMCs were placed in the bottom chamber of the transwell system. Te migratory index was calculated by dividing the number of monocytes that migrated in response to the supernatants from cultured PMCs by the number of monocytes that migrated in response to the control. ∗vs the MO-PBS group, ∗∗P < 0.01. #vs the MO-PMC group, ##P < 0.01 (n 4).

Article Snippet: To investigate whether CR1, CR3, and CXCL12 contributed to monocyte migration, blocking experiments were performed by mixing monocytes with anti-CR1 mAb (5 μg/ml, BM2364, Origene), anti-CR3 mAb (10 μg/ml, 101213, Biolegend) or rabbit IgG irrelevant isotype control.

Techniques: Migration, Staining, Cell Culture, Control