Journal: iScience

Article Title: The Drosophila connectome reveals axo-axonic synapses on descending neurons

doi: 10.1016/j.isci.2026.115624

Figure Lengend Snippet: Axcs make cholinergic axo-axonic synapses on the GFs as predicted by the MANC connectome (A) AN08B098-type neurons (green) form axo-axonic connections at distinct locations along giant fiber (red, GF) axons. The scale bars shown are 20 μm. (B) The AN08B098-GF connections reconstructed in Neuroglancer (Neuroglancer scale 530.45 μm/vh) mirror the morphology seen with fluorescence in A . (C) We validated AN08B098-to-GF connectivity using anti-bruchpilot to stain for T-bars in active zones (white) along AN08B098-type neurons and colocalized the resulting fluorescence with the GFs (red), which were filled with tetramethylrhodamine. The scale bars shown are 20 μm. (D) Neuroglancer reveals presynaptic sites at similar locations seen in colocalized fluorescent image (Neuroglancer scale 530.45 μm/vh). (E and F) XY and XZ plane views of the preparation shown in (A) and (B). The zoomed-in 6 μm (scale bars 3 μm) inlays show AN08B098 forming a single synapse with the GF in (E). The yellow arrows detail the precise location AN08B098 forms the Brp-positive chemical synapse (white) to the GFs in (F). Scale bars shown are 5 μm. (G–O) EM images showing monosynaptic connections between single GF (green) and AN08B098 neurons identified by the following MANC id: (G and H) 21041, (I) 21589, (J) 23949, (K) 152261, (L) 16900, (M) 20444, (N) 22275, and (O) 24038. Pre-and postsynaptic sites are detected in EM slices using a 3D convolutional neural network to identify T-bars (cyan dots) and postsynaptic densities (PSDs, magenta dots). (P–S) We expressed anti-choline acetyltransferase (anti-ChAT) and anti-GFP in AN08B098 neurons. Anti-ChAT colocalizes to AN08B098 cells, with particularly bright staining in the cell bodies. This finding suggests acetylcholine synthesis is present within these cells, validating connectome transmitter predictions for AN08B098. Scale bars shown are (P) 20 μm and (Q–S) 5 μm.

Article Snippet: A mouse primary antibody against bruchpilot (1:50, DSHB, NC82) was used to label presynaptic chemical active zones and coupled to secondary Goat anti-mouse Alexa Fluor 647 (1:500, 115-605-003).

Techniques: Fluorescence, Staining

Journal: Hepatology Communications

Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

doi: 10.1097/HC9.0000000000000928

Figure Lengend Snippet: CCR7 + (c4) MoMFs drive MASLD progression through enhanced antigen presentation and are recruited via the CCL19/CCL21 axis. (A) Feature plots displaying the expression of H2-Eb2 , Ccr7 , Cd209a , and Ciita . (B) Flow cytometry analysis showing CCR7 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (C) Correlation between the proportion of CCR7 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D, E) Dot plots show the enriched GO (D) and KEGG (E) terms of DEGs in CCR7 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (F) Dot plots show the average expression levels of selected DEGs. Dot size denotes the percentage of expressed cells, and the color bar denotes scaled average expression. (G) Flow cytometry analysis showing MHC-II expression levels in CCR7 + MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (H) Percentage of CCR7 + cells among total intrahepatic MoMFs in mice. (I, J) Plasma ALT and AST levels in each group. (K–M) Representative images (K) and quantification of H&E staining (L) and Sirius red staining (M) in liver paraffin sections. Scale bars, 200 μm. (N) Relative mRNA levels of Ccl19 (left) and Ccl21 (right) in liver tissues of NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (O) Feature plots displaying the expression of Top2a , Mki67 , and Stmn1 . (P) Flow cytometry analysis showing Ki67 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (Q) Dot plots show the enriched GO terms of DEGs in Ki67 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. (R) Feature plots displaying the expression of Cd3d , Gzma , and Gzmb . (S) Flow cytometry analysis showing CD3 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (left) and quantitative data (right). (T) Dot plots show the enriched GO terms of DEGs in CD3 + MoMFs between the NCD and MCD groups. Dot size denotes the enriched gene counts. The color bar denotes the adjusted p -value of enrichment. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; H&E, Hematoxylin and Eosin; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

Article Snippet: To further determine the functional contribution of CCR7 + MoMFs, we administered a neutralizing antibody against CCR7 (MAB3477, R&D Systems) or an IgG2A isotype (clone 4B12, R&D Systems) to MCD-fed mice.

Techniques: Immunopeptidomics, Expressing, Flow Cytometry, Activity Assay, Clinical Proteomics, Staining, Derivative Assay, Control

Journal: Hepatology Communications

Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

doi: 10.1097/HC9.0000000000000928

Figure Lengend Snippet: The basal cluster 1 (c1) is depleted during MASLD progression and acquires CD14 + and CCR7 + phenotypes under inflammatory stimulation in vitro. (A) Feature plots displaying the expression of Uba52 , Rpl27 , Rpl15 , and Ifi27l2a . (B) The typical flow cytometry gating strategy and statistical analysis of cluster 1 relative to total MoMFs in NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (C) Correlation between the proportion of cluster 1 and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D) UMAP plots displaying the major cellular subsets identified in the NCD and MCD groups. (E) Trajectory analysis of clusters 0, 1, 4, and 5 using Monocle 2. (F) Heatmap displaying gene expression dynamics across pseudotime for selected marker genes in clusters 0, 1, 4, and 5. (G–I) Flow cytometry analysis of CD14 (G), CCR7 (H), and CCR3 (I) expression in cluster 1 cells isolated from NCD-fed mouse liver, with or without LPS stimulation. (J) Dot plots depicting the expression of genes in liver macrophages derived from healthy and MASLD samples ( GSE212837 ). Abbreviations: CDHFD, choline-deficient high-fat diet; HFD, high-fat diet; LPS, lipopolysaccharide; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; NCD, normal control diet; UMAP, uniform manifold approximation and projection.

Article Snippet: To further determine the functional contribution of CCR7 + MoMFs, we administered a neutralizing antibody against CCR7 (MAB3477, R&D Systems) or an IgG2A isotype (clone 4B12, R&D Systems) to MCD-fed mice.

Techniques: In Vitro, Expressing, Flow Cytometry, Activity Assay, Gene Expression, Marker, Isolation, Derivative Assay, Control

Journal: Hepatology Communications

Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

doi: 10.1097/HC9.0000000000000928

Figure Lengend Snippet: Heterogeneity and dynamics of intrahepatic MoMFs during MASLD progression. This scRNA-seq analysis delineated 7 distinct MoMFs clusters (c0–c6) in the liver with dynamic changes during steatohepatitis progression. CD14 + (c0) MoMFs, the predominant CCR2 + population, exhibited multifaceted functions beyond inflammation, including roles in fibrosis, lipid uptake, and antigen presentation. The basal cluster (c1) is depleted during MASLD progression and acquires CD14 + and CCR7⁺ phenotypes under inflammatory stimulation in vitro. Lipid overload drives mitochondrial dysfunction and apoptosis in CD206 + (c2) MoMFs in MASLD. CCR3 + (c3) MoMFs drive MASLD progression through lipid accumulation, oxidative stress, and CCL5-mediated recruitment. CCL19/CCL21-recruited CCR7 + (c4) MoMFs exacerbate MASLD pathogenesis through enhanced antigen presentation. In addition, Ki67 + (c5) proliferative and CD3 + (c6) TCR-related MoMFs maintained stable proportions but exhibited functional alterations during MASLD. Abbreviations: MASLD, metabolic dysfunction–associated steatotic liver disease; MoMFs, monocyte-derived macrophages; scRNA-seq, single-cell RNA sequencing.

Article Snippet: To further determine the functional contribution of CCR7 + MoMFs, we administered a neutralizing antibody against CCR7 (MAB3477, R&D Systems) or an IgG2A isotype (clone 4B12, R&D Systems) to MCD-fed mice.

Techniques: Immunopeptidomics, In Vitro, Functional Assay, Derivative Assay, Single Cell, RNA Sequencing

Journal: Genes & Diseases

Article Title: Chloride channel accessory 4 suppresses stem cell-like properties of colorectal cancer and enhances anti-PD-1 immunotherapy

doi: 10.1016/j.gendis.2025.101859

Figure Lengend Snippet: CLCA4 overexpression enhanced the therapeutic effect of anti-PD-1. (A) In vivo tumorigenicity assay in nude mice was performed to detect the therapeutic effect of CLCA4 overexpression combined with anti-PD-1 antibody. (B) Tumor growth curve was monitored from nude mice with different treatment groups. (C) Weights of tumors from nude mice with different treatments were detected. One-way ANOVA with Tukey's multiple comparisons test (mean ± standard deviation). (D) Analysis of the survival times of mice in each group ( n = 8 per group), and the experiment was terminated 60 days after tumor inoculation. Unpaired two-tailed t -test (mean ± standard deviation). (E) Immunofluorescence or immunohistochemistry staining was performed to detect the infiltration levels of CD8 + T cells, GZMB, Perforin + cells, Ki67 + , Bmi-1 + , and Oct4 + cells in tumors from different treatment groups. One-way ANOVA with Tukey's multiple comparisons test (mean ± standard deviation). (F) Hematoxylin-eosin staining of liver metastases in each group. (G) Quantitative analysis of the liver weight in each group. One-way ANOVA with Tukey's multiple comparisons test (mean ± standard deviation). (H) Quantitative analysis of the liver metastasis area/total liver area in each group. One-way ANOVA with Tukey's multiple comparisons test (mean ± standard deviation).

Article Snippet: After 7 days, mice were intraperitoneally treated with either an in vivo blocking antibody against mouse PD-1 (Clone: 29F.1A2, BioXcell, Cat# BP0273) or a rat IgG2a isotype control antibody (Clone: 2A3, BioXcell, Cat# BP0089).

Techniques: Over Expression, In Vivo, Tumorigenicity Assay, Standard Deviation, Two Tailed Test, Immunofluorescence, Immunohistochemistry, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: BPGM as an intrinsic brake to constrain metastasis through phospho-epigenetic-mediated carnitine biosynthesis suppression

doi: 10.1016/j.neo.2026.101299

Figure Lengend Snippet: Overexpression of BPGM inhibits tumor metastasis in vitro and in vivo . (A) Silencing BPGM promoted migration of tumor cells. Tumor cells stably expressing shBPGM and its control cells (shCtrl) were examined. Scale bar, 200 μm. (B) Overexpressing BPGM suppressed migration of tumor cells. Tumor cells stably expressing BPGM and its control cells (Ctrl) were examined. Scale bar, 200 μm. (C) The mRNA levels of MMP2 and MMP9 reduced in BPGM-overexpressing tumor cells. SK-HEP-1 cells stably expressing BPGM (BPGM-OE) and its ctrl cells were employed to detect the mRNA levels of MMP2 and MMP9 by qPCR analysis. β-actin was used as an internal control. (D-E) Xenografts of stable Bpgm-overexpressing cells displayed a lower rate of liver and lung metastasis and less metastatic nodules in the liver. For (D), Hepa-Ctrl ( n = 7) and Hepa-BPGM sublines ( n = 6) were inoculated under the capsule of the left hepatic lobe of C57BL/6 mice. Upper panel, a schematic diagram of orthotopic hepatic implantation model. The number of metastatic rate and nodules is shown (D, lower panel). Scale bar, 1 cm. Hematoxylin-eosin staining was performed on serial sections of livers (E, left panel) and lungs (E, right panel) to detect the metastatic nodules. The red arrows indicated the metastatic nodules. Scale bar in left panel, 200 µm; Scale bar in right panel, 100 μm. (F-H) Overexpressing of BPGM inhibited lung metastasis of tumor xenografts. Scale bar in F, 1 cm. B16-F10 cells transfected with Ctrl ( n = 6) or BPGM-OE ( n = 6) was injected into the tail vein of C57BL/6 mice, respectively. Upper panel in F, a schematic diagram of lung metastasis model by tail vein injection. H&E staining of lung sections was performed to observe metastatic foci (G). For G, scale bar in left panel, 500 μm; scale bar in right panel, 200 μm. The number of melanoma nodules and lung metastasis is shown in H. (I) The model deciphers the inhibitory role of BPGM in tumor metastasis. Error bar: mean ± SEM. P -values are labeled above the bar chart.

Article Snippet: The antibodies used included mouse antibody against β-actin (BM0627, Boster, Wuhan, China), rabbit antibody against BPGM (17173-1-AP, Proteintech), EZH2 (F0281, Selleck), phospho-EZH2 (Thr345) (TA3584S, Abmart, Shanghai, China), phospho-CDK1 (Thr14) (AP1465, Abclonal, Wuhan, China), ubiquitin (10201-2-AP, Proteintech), HIF1α (36169, Cell Signaling Technology, CST, Beverly, MA, USA), H3K4me3 (91264, Active Motif), H3K79me3 (cat 49-1020, Thermos Fisher), H3K9me3 (61014, Active Motif), H3K27me3 (91168, Active Motif) and Histone 3 (F0057, Selleck).

Techniques: Over Expression, In Vitro, In Vivo, Migration, Stable Transfection, Expressing, Control, Staining, Transfection, Injection, Labeling