Journal: Advanced Science

Article Title: Aged ZnO Nanoparticles Promote Breast Cancer Lung Metastasis by Remodeling the Pulmonary Microenvironment Through the CXCL10/Wnt/β‐Catenin Axis

doi: 10.1002/advs.78249

Figure Lengend Snippet: CXCL10 silencing abrogates aged ZnO NPs–induced accumulation of pro‐metastatic stromal populations. (A) Schematic of coculture model: NIH‐3T3 or Raw264.7 cells with CXCL10 knockdown were stimulated with PBS, fresh or aged NPs (7.5 µg/mL), co‐cultured with 4T1 cells for 72 h. (B) qPCR analysis confirming CXCL10 knockdown in Raw264.7 cells ( n = 3). (C‐E) CXCL10 knockdown in Raw264.7 macrophages attenuates ZnO NP‐enhanced M2 polarization, as shown by reduced Arg1, Mrc1 expression (C) and CD206 + cell frequency (D, E) ( n = 3). (F) qPCR analysis confirming CXCL10 knockdown in NIH‐3T3 cells ( n = 3). (G, H) Flow cytometric analysis (G) and quantification (H) of the percentage of CD140a‐positive cells among CXCL10‐knockdown NIH‐3T3 cells in the co‐culture system ( n = 3). (I) qPCR analysis of CAF‐related genes (FN1, FSP1, α‐SMA, CD140a) in CXCL10‐knockdown NIH‐3T3 cells from the co‐culture model ( n = 3). (J) Schematic illustration of primary MDSCs preparation: primary mouse bone marrow‐derived macrophages were differentiated into MDSCs by supplement of GM‐CSF (40 ng/mL) and IL‐6 (40 ng/mL). MDSCs with CXCL10 knockdown were stimulated with PBS, fresh ZnO NPs, or aged ZnO NPs (7.5 µg/mL), then co‐cultured with 4T1 cells for 72 h. Cells were collected for further experiments. (K) qPCR analysis of relative CXCL10 mRNA expression in MDSCs with CXCL10 knockdown ( n = 3). (L, M) Flow cytometric analysis (L) and quantification (M) of the proportion of granulocytic CD11b + Ly6G + Ly6C low PMN‐MDSCs in the co‐culture system after CXCL10 knockdown ( n = 3). All data were represented as the mean ± SD. P values were calculated by one‐way ANOVA followed by Tukey's test. * p <0.05; ** p <0.01.

Article Snippet: Sections were then incubated overnight at 4°C with the following primary antibodies: F4/80 (Servicebio, GB11027; 1:750), Ki‐67 (Abcam, ab15580; 1:200), FSP‐1 (Bioss, bs‐1015R; 1:250), FN‐1 (BD Pharmingen, 610077; 1:250), CD206 (Affinity, DF7564; 1:250), αSMA (Affinity, AF1032; 1:500), bFGF (Proteintech, 14726‐1‐AP; 1:200), COX‐2 (Cell Signaling Technology, 12282S; 1:100).

Techniques: Knockdown, Cell Culture, Analysis, Expressing, Co-culture Assay, Derivative Assay

Journal: bioRxiv

Article Title: A Conserved Chromatin-Driven Checkpoint Defines Late Macrophage Maturation Independent of Tissue Specialization

doi: 10.64898/2026.07.01.735834

Figure Lengend Snippet: A) Genomic Locus Overlap Enrichment Analysis (LOLA) of differentially accessible regions (DARs) between day 13 and day 11, stratified according to the four clusters defined in A) (open promoter, n = 477 DARs; open distal, n = 368 DARs; closed promoter, n = 309 DARs; closed distal, n = 1,008 DARs). Enrichment was performed against the database of transcriptional regulators peaks from ReMap2022. Statistical significance was assessed using Fisher’s exact test implemented in LOLA (FDR-adjusted p < 0.05). The x-axis represents the odds ratio, and adjusted FDR values are indicated in the labels (ns, grey; FDR < 0.05, light blue; FDR < 0.005, blue; FDR < 0,0005 dark blue). The background universe corresponds to all accessible regions detected in the ATAC-Seq dataset. B) Functional enrichment analysis of the nearest associated genes to the defined DARs using ChEA gene sets. , Enrichment was assessed using a one-sided hypergeometric test with false discovery rate correction (Benjamini–Hochberg). The x-axis shows the number of enriched genes, and adjusted P values are indicated in the labels. The background gene universe corresponds to all genes associated with peak detected in the ATAC-seq dataset. C) Heatmap of differentially expressed genes (DEGs) induced by BAF inhibition, identified by comparing BAF inhibitor–treated samples (BRM/BRG1 ATP Inhibitor-1 - BRM014; 0.016, 0.08, and 0.4 µM; pink gradient from light to dark) to untreated controls. The inhibitor was applied at day 12 and RNA was collected at day 13 (DESeq2 with apeglm LFC shrinkage; adjusted p < 0.05; |fold change| > 1.5). Expression values are scaled by row (turquoise to yellow).

Article Snippet: On day 12, cells were treated for 24 h with the BRM/BRG1 ATPase (SWI/SNF) inhibitor BRM014 (MedChemExpress, cat. no. HY-119374) at final concentrations of 400, 80, or 16 nM, with PBS-treated wells included as controls.

Techniques: Functional Assay, Inhibition, Expressing