Journal: Journal of Inflammation Research

Article Title: Comprehensive Gene Analysis Reveals Cuproptosis-Related Gene Signature Associated with M2 Macrophage in Staphylococcus aureus -Infected Osteomyelitis

doi: 10.2147/JIR.S457414

Figure Lengend Snippet: Immune microenvironment analysis and M2 macrophage-related CRGs (M2R-CRGs) selection. ( A ) Immune landscape of 22 immune cell types in osteomyelitis patients. ( B ) Significant differences in the infiltration levels of 12 immune cell types between osteomyelitis patients and healthy controls. ( C ) Correlation analysis of DE-CRGs with 22 immune cell types to select M2R-CRGs. ( D ) Venn diagram of M2R-CRGs. ( E ) Differences in the infiltration abundance of 22 immune cell types between the high expression group and low expression group of SLC31A1. ( F ) Differences in the infiltration abundance of 22 immune cell types between the high expression group and low expression group of DLD. ( G ) Differences in the infiltration abundance of 22 immune cell types between the high expression group and low expression group of MTF1. Significance: * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The primary antibodies used were as follows: SLC31A1 (DF13356; Affinity Biosciences, Suzhou, China, 1:100), DLD (DF7322; Affinity Biosciences, Suzhou, China, 1:100), MTF1 (Bs-3601R; Bioss Antibodies, Beijing, China, 1:100)。

Techniques: Selection, Expressing

Journal: Journal of Inflammation Research

Article Title: Comprehensive Gene Analysis Reveals Cuproptosis-Related Gene Signature Associated with M2 Macrophage in Staphylococcus aureus -Infected Osteomyelitis

doi: 10.2147/JIR.S457414

Figure Lengend Snippet: Single-gene GSEA-KEGG pathway analysis in SLC31A1 ( A ), DLD ( B ), MTF1 ( C ). Single-gene GSVA-KEGG pathway analysis in SLC31A1 ( D ), DLD ( E ), MTF1 ( F ).

Article Snippet: The primary antibodies used were as follows: SLC31A1 (DF13356; Affinity Biosciences, Suzhou, China, 1:100), DLD (DF7322; Affinity Biosciences, Suzhou, China, 1:100), MTF1 (Bs-3601R; Bioss Antibodies, Beijing, China, 1:100)。

Techniques:

Journal: Journal of Inflammation Research

Article Title: Comprehensive Gene Analysis Reveals Cuproptosis-Related Gene Signature Associated with M2 Macrophage in Staphylococcus aureus -Infected Osteomyelitis

doi: 10.2147/JIR.S457414

Figure Lengend Snippet: Validation for 3 M2R-CRGs through independent dataset (GSE30119), RT-qPCR, and immunohistochemistry. In the independent dataset, SLC31A1 ( A ), DLD ( B ), and MTF1 ( C ) were all significantly upregulated in the osteomyelitis rat group. Total RNA and protein were extracted from both the osteomyelitis rat group and the sham operation group. The mRNA expression levels of SLC31A1 ( D ), DLD ( E ), and MTF1 ( F ) were significantly increased in the osteomyelitis rat group. Furthermore, the protein levels of SLC31A1 ( G ), DLD ( H ), and MTF1 ( I ) showed a significant upregulation in the osteomyelitis rat group. Significance: *: P <0.05, **: P <0.01, ***: P <0.001, ****: P <0.0001.

Article Snippet: The primary antibodies used were as follows: SLC31A1 (DF13356; Affinity Biosciences, Suzhou, China, 1:100), DLD (DF7322; Affinity Biosciences, Suzhou, China, 1:100), MTF1 (Bs-3601R; Bioss Antibodies, Beijing, China, 1:100)。

Techniques: Quantitative RT-PCR, Immunohistochemistry, Expressing

Journal: bioRxiv

Article Title: The PBAF chromatin remodeling complex contributes to metal homeostasis through Mtf1 regulation

doi: 10.1101/2025.04.12.648552

Figure Lengend Snippet: Representative confocal microscopy images showing the expression and nuclear localization of Baf180 (red) and Mtf1 (green) in wild type (A) , scr (B) , Baf180 KD (C) , Baf250a KD (D) and Brd9 KD (E) C2C12 myoblasts. Nuclei are counterstained with DAPI (blue). (F) Quantification of Mtf1 colocalization with Baf180 in control and the different SWI/SNF KD myoblasts from panel A-E and from supplemental figures 6 and 7. Colocalization is disrupted in Baf180 KD cells, while metal treatment modulates Mtf1-Baf180 interactions. Scale bars: 10 μm (panoramic views), 5 μm (zoomed-in images). Mean ± SE of three independent biological replicates. *P < 0.05, **P < 0.01, ****P < 0.0001. * Indicates significance relative to untreated WT cells. (G) Representative immunoprecipitation (IP) of Mtf1 or Baf180 in proliferating wild type myoblasts (N=3). Input, a Baf180 IP obtained from Baf180 KD cells and an IgG IP were included as controls. Mtf1 exhibits significantly higher colocalization with Baf180 compared to Baf250a or Brd9 in WT proliferating myoblasts. To show both proteins, the western blot was developed using both the rabbit anti-Baf180 and the mouse anti-Mtf1 specific antibodies, incubated sequentially in the same membrane, followed by sequential incubation of species-specific secondary antibodies.

Article Snippet: Normal rabbit IgG (sc-2027) and mouse anti-Mtf1 (sc-365090) were obtained from Santa Cruz Biotechnologies.

Techniques: Confocal Microscopy, Expressing, Control, Immunoprecipitation, Western Blot, Incubation, Membrane

Journal: bioRxiv

Article Title: The PBAF chromatin remodeling complex contributes to metal homeostasis through Mtf1 regulation

doi: 10.1101/2025.04.12.648552

Figure Lengend Snippet: CUT&RUN analysis illustrating Mtf1 binding dynamics in proliferating myoblasts under untreated conditions and following 100 μM CuSO 4 supplementation for 48 h. (A) Combined CUT&RUN heatmaps from peak calling of three independent biological sequences obtained for each condition. Normal IgG used as control. (B) Aggregation plots of Mtf1 CUT&RUN data showing occupancy of Mtf1 and IgG over TSS’s in the presence or absence of Cu. (C) Overlap of CUT&RUN peaks of Mtf1 across the genome observed in proliferating cells in the presence or absence of Cu. See complete set of genes in Supp. Table 5 . (D) Pie charts showing the binding site locations for Mtf1 in myoblasts cultured in the presence or absence of Cu and combined in both conditions. (E) Main changes of Mtf1 motif-binding dependent on Cu supplementation in proliferating C2C12 myoblasts myoblasts. Novel consensus DNA-binding motifs identified from peak calling for CUT&RUN within Mtf1 peaks in proliferating cells supplemented or not with Cu and in both conditions combined. The top five most significant motifs enriched, including the DNA logo, its corresponding transcription factor, and its P value are shown. (F) GO analysis of CUT&RUN data showing the main categories of genes that Mtf1 binds to in proliferating myoblasts in both the absence or presence of 100 μM CuSO 4 and in both conditions combined. Cut-off was set at 2.0 of the -log(adjusted P value). Consensus DNA-binding motifs within Mtf1 peaks reveal distinct sequence preferences in Cu-treated versus untreated cells.

Article Snippet: Normal rabbit IgG (sc-2027) and mouse anti-Mtf1 (sc-365090) were obtained from Santa Cruz Biotechnologies.

Techniques: Binding Assay, Control, Cell Culture, Sequencing