Journal: Current Issues in Molecular Biology

Article Title: MSTN and TCF12 as Candidate Immunometabolic Signatures in Glioma-Associated Foam Cells: Insights from Integrated Multi-Omics Analysis

doi: 10.3390/cimb48030289

Figure Lengend Snippet: Identification and immune correlates of prognostic genes in TAFCs. ( A ) RSF analysis of TAFC-specific markers identifies three genes ( MSTN , TCF12 , and RPL3 ) as top candidates, with values showing variable relative importance > 0.4. ( B , C ) Kaplan–Meier survival curves showing that high expression of MSTN or TCF12 is associated with significantly poorer overall survival in glioma patients. The median survival probability differs significantly between the high-expression and low-expression groups. (log-rank test, MSTN : p = 0.0012; TCF12 : p = 0.0029), whereas RPL3 showed no prognostic relevance. ( D ) Percentage of immune cells in control (green) and glioma (purple) groups. ( E ) Interaction analysis of 22 different immune cells in glioma patients (blue: negative correlation; red: positive correlation). ( F ) Comparisons of immune cells in control (blue) and glioma (purple) groups (*: p < 0.05; **: p < 0.01; ***: p < 0.001; ns: not significant). ( G ) Correlation analysis demonstrates that MSTN expression positively correlates with M2 macrophages (correlation > 0.21, p < 0.01) and γδ T cells (correlation = 0.17, p < 0.05), and negatively correlates with M0 macrophages (correlation = −0.21, p < 0.05) and Tregs (correlation = −0.19, p < 0.05). ( H ) TCF12 expression shows a positive correlation with M2 macrophages (correlation = 0.27, p < 0.005) and resting mast cells (correlation = 0.18, p < 0.05), and a negative correlation with activated mast cells (correlation = −0.24, p < 0.005) and monocytes (correlation = −0.27, p < 0.005).

Article Snippet: After blocking endogenous peroxidase activity with 3% H 2 O 2 and non-specific sites with 3% BSA, the sections were incubated overnight at 4 °C with the following primary antibodies: TCF12 (1:200; Proteintech, 14419-1-AP, Wuhan, China), MSTN (1:200; Proteintech, 19142-1-AP, Wuhan, China), and CD163 (1:200; Proteintech, 68218-1-ig, Wuhan, China).

Techniques: Expressing, Control

Journal: Current Issues in Molecular Biology

Article Title: MSTN and TCF12 as Candidate Immunometabolic Signatures in Glioma-Associated Foam Cells: Insights from Integrated Multi-Omics Analysis

doi: 10.3390/cimb48030289

Figure Lengend Snippet: Associations of MSTN and TCF12 with disease-related genes and immune–metabolic pathway activities. ( A ) Expression profiles of the top 20 glioma-related genes (ranked by transcription levels from GeneCards), highlighting differentially expressed genes such as LGI1, POT1, and H19 (blue: control group; yellow: tumor group). ( B ) Bubble map for the Pearson correlations between MSTN and TCF12 and disease-related genes. Larger circles indicate that the p -value is closer to zero; a redder color indicates a stronger positive correlation and a deeper purple indicates a stronger negative correlation. ( C ) Single-cell pathway activity scores indicating elevated activity of MSTN and TCF12 in immune–metabolic pathways, including cholesterol homeostasis, androgen response, and angiogenesis. A larger dot size indicates a higher enrichment significance (FDR closer to 0); logFC represents fold change; red indicates a positive correlation, and blue indicates a negative correlation. ( D ) Single-cell metabolic pathway activity associated with MSTN and TCF12 expression. Y -axis: 5 major metabolic signatures including amino acid, lipid, drug, other metabolism, and C3-specific pathways. Red indicates high pathway activity; blue indicates low pathway activity. X -axis: yellow = high gene expression group; cyan = low gene expression group. *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001.

Article Snippet: After blocking endogenous peroxidase activity with 3% H 2 O 2 and non-specific sites with 3% BSA, the sections were incubated overnight at 4 °C with the following primary antibodies: TCF12 (1:200; Proteintech, 14419-1-AP, Wuhan, China), MSTN (1:200; Proteintech, 19142-1-AP, Wuhan, China), and CD163 (1:200; Proteintech, 68218-1-ig, Wuhan, China).

Techniques: Expressing, Control, Single Cell, Activity Assay, Gene Expression

Journal: Current Issues in Molecular Biology

Article Title: MSTN and TCF12 as Candidate Immunometabolic Signatures in Glioma-Associated Foam Cells: Insights from Integrated Multi-Omics Analysis

doi: 10.3390/cimb48030289

Figure Lengend Snippet: Transcriptional and post-transcriptional regulatory networks of MSTN and TCF12 , and their association with functional pathways. ( A ) miRNA–mRNA interaction network predicted by the MiRcode database showing 63 miRNAs that could potentially regulate MSTN and TCF12 . Key miRNAs such as miR-129-5p, miR-219-5p, and miR-221 are highlighted. ( B ) Transcriptional regulatory network illustrating TFs enriched in the promoters of MSTN and TCF12 . The most significantly enriched motif is cisbp_M1404 (NES = 5.82). ( C – F ) GSEA reveals pathways significantly associated with MSTN (ribosome, oxidative phosphorylation, Fanconi anemia) and TCF12 (mRNA surveillance, Notch signaling, Fanconi anemia). ( G , H ) GSVA highlights distinct functional profiles: MSTN correlates with DNA repair and spermatogenesis pathways, while TCF12 is associated with G2/M checkpoint and E2F target pathways.

Article Snippet: After blocking endogenous peroxidase activity with 3% H 2 O 2 and non-specific sites with 3% BSA, the sections were incubated overnight at 4 °C with the following primary antibodies: TCF12 (1:200; Proteintech, 14419-1-AP, Wuhan, China), MSTN (1:200; Proteintech, 19142-1-AP, Wuhan, China), and CD163 (1:200; Proteintech, 68218-1-ig, Wuhan, China).

Techniques: Functional Assay, Phospho-proteomics

Journal: Current Issues in Molecular Biology

Article Title: MSTN and TCF12 as Candidate Immunometabolic Signatures in Glioma-Associated Foam Cells: Insights from Integrated Multi-Omics Analysis

doi: 10.3390/cimb48030289

Figure Lengend Snippet: Spatial heterogeneity and differentiation-associated expression dynamics of MSTN and TCF12 . ( A , B ) Spatial distribution of MSTN and TCF12 expression in two independent glioma samples ( GSM8340238 and GSM8340239 ) from the GEO database. Color scale represents normalized MSTN and TCF12 expression levels, with blue indicating low expression and red indicating high expression. ( C ) Pseudotime trajectories displayed in component space. The color gradient from dark blue to light yellow represents the pseudotime value, with dark blue indicating the start of the trajectory (early differentiation) and light yellow indicating the end (late differentiation). ( D ) Distribution of cell states (State 1, 2, 3) in component space, with distinct colors representing different cell states. ( E ) Distribution of cell clusters (Clusters 0–6) in component space, with distinct colors representing seven different cell clusters. ( F , G ) Projection of pseudotime trajectories onto corresponding HE-stained sections enables dual-dimensional “pseudotime-spatial” visualization, with different colors representing the clustered cell clusters. ( H ) Expression dynamics of MSTN and TCF12 along the pseudotime axis. Each colored line represents the relative expression level of MSTN (left) and TCF12 (right) for a specific cell type, with colors corresponding to the cell types defined in ( J ). ( I ) Expression dynamics of MSTN and TCF12 along the pseudotime axis across three cell states (State 1, 2, 3). The color gradient from purple to yellow represents the pseudotime value, with purple indicating early pseudotime and yellow indicating late pseudotime. Each point represents a cell, colored by its state: State 1 (red), State 2 (blue), and State 3 (green). ( J ) Cell type annotation legend showing the eight major cell types identified, with distinct colors corresponding to each cell type: Tumor associated foam cell (red), Excitatory neuron (blue), Oligodendrocyte precursor cell (green), Inhibitory neuron (purple), Microglial (orange), Astrocyte (yellow), Endothelial cell (cyan), and Oligodendrocyte (pink).

Article Snippet: After blocking endogenous peroxidase activity with 3% H 2 O 2 and non-specific sites with 3% BSA, the sections were incubated overnight at 4 °C with the following primary antibodies: TCF12 (1:200; Proteintech, 14419-1-AP, Wuhan, China), MSTN (1:200; Proteintech, 19142-1-AP, Wuhan, China), and CD163 (1:200; Proteintech, 68218-1-ig, Wuhan, China).

Techniques: Expressing, Staining

Journal: Current Issues in Molecular Biology

Article Title: MSTN and TCF12 as Candidate Immunometabolic Signatures in Glioma-Associated Foam Cells: Insights from Integrated Multi-Omics Analysis

doi: 10.3390/cimb48030289

Figure Lengend Snippet: Association of MSTN and TCF12 with chemotherapy sensitivity and a clinical prognostic nomogram. ( A , B ) Correlation analysis between expression of MSTN or TCF12 and predicted sensitivity to common chemotherapeutic agents from the GDSC database. MSTN expression correlates with sensitivity to paclitaxel ( p = 0.0011) and VE-822 ( p = 0.048), while TCF12 expression is associated with sensitivity to cytarabine ( p = 0.00031), olaparib ( p = 1.8 × 10 −8 ), a WEE1 inhibitor ( p = 3 × 10 −4 ), paclitaxel ( p = 0.00041), and VE-822 ( p = 1.5 × 10 −6 ). ( C ) Prognostic nomogram integrating MSTN and TCF12 expression with clinical variables (age, gender, chemotherapy regimen). ( D ) Nomogram calibrated for 2- and 3-year overall survival prediction for TCGA glioma patients. ( E ) Decision curve analysis of prediction models (net benefit across risk thresholds (0–1)). Blue: combined model; red: MSTN ; yellow: TCF12 ; green: age; cyan: gender; gray: all. The combined model demonstrates the greatest clinical utility in the 0.3–0.7 threshold range.

Article Snippet: After blocking endogenous peroxidase activity with 3% H 2 O 2 and non-specific sites with 3% BSA, the sections were incubated overnight at 4 °C with the following primary antibodies: TCF12 (1:200; Proteintech, 14419-1-AP, Wuhan, China), MSTN (1:200; Proteintech, 19142-1-AP, Wuhan, China), and CD163 (1:200; Proteintech, 68218-1-ig, Wuhan, China).

Techniques: Expressing

Journal: Current Issues in Molecular Biology

Article Title: MSTN and TCF12 as Candidate Immunometabolic Signatures in Glioma-Associated Foam Cells: Insights from Integrated Multi-Omics Analysis

doi: 10.3390/cimb48030289

Figure Lengend Snippet: Histological and immunofluorescence validation of MSTN and TCF12 expression in TAFC-enriched glioma tissues. ( A ) Representative HE staining of paired peritumoral and glioma tissues. Peritumoral regions show an abundance of microglia, while tumor areas are enriched with foam-like cells The two smaller panels below are magnified views of the areas outlined by the red boxes. Red arrows indicate macrophages.. ( B , C ) Immunofluorescence staining confirms significant upregulation of MSTN ( B ) and TCF12 ( C ) in glioma compared to adjacent tissues. ( D ) Quantitative analysis of MSTN and TCF12 fluorescence intensity. ( E , F ) Dual-color immunofluorescence co-staining of macrophage marker CD163 and MSTN ( E ) and TCF12 ( F ). Yellow signal indicates co-localization, which is markedly increased in tumor tissues. The right panel shows a magnified view of the blue box. ( G ) Quantification of the percentage of CD163 + macrophages co-positive for MSTN or TCF12. Nuclei are counterstained with DAPI. Green: CD163; red: MSTN or TCF12; blue: DAPI. Data is presented as mean ± SEM ( n = 5). Scale bars = 50 μm. *: p < 0.05; **: p < 0.01.

Article Snippet: After blocking endogenous peroxidase activity with 3% H 2 O 2 and non-specific sites with 3% BSA, the sections were incubated overnight at 4 °C with the following primary antibodies: TCF12 (1:200; Proteintech, 14419-1-AP, Wuhan, China), MSTN (1:200; Proteintech, 19142-1-AP, Wuhan, China), and CD163 (1:200; Proteintech, 68218-1-ig, Wuhan, China).

Techniques: Immunofluorescence, Biomarker Discovery, Expressing, Staining, Fluorescence, Marker

Journal: bioRxiv

Article Title: Pathological TDP-43 filaments accumulate at synapses and cause synaptic dysfunction

doi: 10.64898/2026.01.27.701787

Figure Lengend Snippet: A. Schematic summarising the extraction of TDP-43 filaments and their incubation with neuronal cultures. B. Immunogold negative-stain electron microscopy of extracted TDP-43 filaments from FTLD-TDP Type A patient brain, using a primary antibody against the N-terminus of TDP-43 and a secondary antibody conjugated to 10 nm gold particles. Scale bar, 100 nm. C. Immunoblot of extraction fractions and the final TDP-43 filament sample from FTLD-TDP Type A patient brain, probed with an antibody against pS409/410 TDP-43 (pTDP-43). The arrow indicates full-length (FL) TDP-43 and the bar indicates C-terminal fragments (CTFs) of TDP-43. D. Immunofluorescence confocal microscopy images of mouse primary cortical neurons incubated with (+ filaments) or without (- filaments) FTLD-TDP Type A patient-derived TDP-43 filaments for 7 d. Neurons were labelled using an antibody against class III β-tubulin (βIII-Tub; cyan) and TDP-43 filaments were labelled using the antibody against pTDP-43 (magenta). E. pTDP-43 signal (white) within images masked using the βIII-tubulin signal (yellow). Signal of 0.05-3 μm 2 is shown in cyan. This size filter was used to exclude noise and large extracellular TDP-43 filament accumulations from quantification in (F) . Scale bar, 20 µm. F. Quantification of βIII-tubulin-masked pTDP-43 signal from (E) at different timepoints. The total area of pTDP-43 signal as a percentage of the βIII-tubulin signal mask is plotted. Each data point represents one technical repeat. Data points are colour coded by biological replicate. n = 3 biological replicates. Means +/- SD are shown. A one-way ANOVA with Tukey’s multiple comparison test was performed, ***p<0.001, ****p<0.0001. G. Immunofluorescence confocal microscopy images of mouse primary cortical neurons incubated with FTLD-TDP Type A patient brain-derived TDP-43 filaments for 1 d. TDP-43 filaments were labelled with the antibody against pTDP-43 pre- (yellow) and post- (magenta) detergent permeabilisation. Neurons were labelled with an antibody against class III β-tubulin (βIII-Tub; cyan). Arrows indicate examples of pTDP-43 signal unique to permeabilised neurons. H. pTDP-43 signal (white) within images masked using the βIII-tubulin signal (yellow). Signal of 0.05-3 μm 2 shown in cyan. This size filter was used to exclude noise and large extracellular TDP-43 filament accumulations from quantification in (I) . Scale bars, 5 and 10 μm, as indicated. I. Quantification of βIII-tubulin-masked pTDP-43 signal pre- and post-permeabilisation from (H) . The total area of pTDP-43 signal as a percentage of the βIII-tubulin signal mask is plotted. Each pair of data points represent one technical repeat. n = 3 biological replicates. A paired t-test was performed, **p<0.01.

Article Snippet: The grids were then blotted and incubated with anti-TDP-43 primary antibody (Proteintech, 10782-2-AP; 1:25) diluted in blocking buffer for 3 h at 21°C.

Techniques: Extraction, Incubation, Staining, Electron Microscopy, Western Blot, Immunofluorescence, Confocal Microscopy, Derivative Assay, Comparison

Journal: bioRxiv

Article Title: Pathological TDP-43 filaments accumulate at synapses and cause synaptic dysfunction

doi: 10.64898/2026.01.27.701787

Figure Lengend Snippet: A. Schematic summarising the antibody-targeted proximity labelling strategy. TDP-43 filaments (red) were targeted using a primary antibody against pS409/410 TDP-43 (magenta). Horseradish peroxidase (grey) conjugated to a secondary antibody (green) was then used to catalyse the conversion of biotin-phenol to biotin-phenoxyl radicals (yellow) in the presence of hydrogen peroxide, thereby biotinylating proximal biomolecules. B. Immunofluorescence confocal microscopy images of mouse primary cortical neurons incubated with (+ filaments) or without (- filaments) FTLD-TDP Type A patient brain-derived TDP-43 filaments for 1 d, followed by antibody-targeted proximity labelling. TDP-43 filaments were labelled using an antibody against pS409/410 TDP-43 (pTDP-43; magenta), biotin was labelled using fluorescence-conjugated streptavidin (yellow) and neurons were labelled with an antibody against class III β-tubulin (βIII-Tub; cyan). Scale bars, 10 μm. C. Immunoblots of affinity purified biotinylated proteins from mouse primary cortical neurons (beads, b) and the flowthrough (f), probed with antibodies against biotin (left) and TDP-43 (right). Neurons were incubated with (+ filaments) or without (- filaments) FTLD-TDP Type A patient brain-derived TDP-43 filaments, in the presence or absence (- antibody) of the antibody against pTDP-43, followed by antibody-targeted proximity labelling.

Article Snippet: The grids were then blotted and incubated with anti-TDP-43 primary antibody (Proteintech, 10782-2-AP; 1:25) diluted in blocking buffer for 3 h at 21°C.

Techniques: Immunofluorescence, Confocal Microscopy, Incubation, Derivative Assay, Fluorescence, Western Blot, Affinity Purification

Journal: bioRxiv

Article Title: Pathological TDP-43 filaments accumulate at synapses and cause synaptic dysfunction

doi: 10.64898/2026.01.27.701787

Figure Lengend Snippet: A. Venn diagrams of the number of significantly-enriched proximal proteins from mouse primary cortical neurons and human ESC-derived cortical neurons incubated with FTLD-TDP Type A patient-brain derived TDP-43 filaments for 1 d and 3 d. B. Bar plot of numbers of significantly-enriched Gene Ontology (GO) Biological Process terms for proximal proteins from mouse primary cortical neurons and human ESC-derived cortical neurons incubated with FTLD-TDP Type A patient-brain derived TDP-43 filaments for 1 d and 3 d. C. Schematic summarising the significantly-enriched GO Cellular Compartment terms (red) for mouse primary cortical neurons and human ESC-derived cortical neurons incubated with FTLD-TDP Type A patient-brain derived TDP-43 filaments for 1 d and 3 d. D. Representative significantly-enriched proximal proteins from mouse primary cortical neurons and human ESC-derived cortical neurons incubated with FTLD-TDP Type A patient-brain derived TDP-43 filaments for 1 d and 3 d.

Article Snippet: The grids were then blotted and incubated with anti-TDP-43 primary antibody (Proteintech, 10782-2-AP; 1:25) diluted in blocking buffer for 3 h at 21°C.

Techniques: Derivative Assay, Incubation

Journal: bioRxiv

Article Title: Pathological TDP-43 filaments accumulate at synapses and cause synaptic dysfunction

doi: 10.64898/2026.01.27.701787

Figure Lengend Snippet: A. Representative significantly-enriched proximal pre- and postsynaptic proteins from mouse primary cortical neurons and human ESC-derived cortical neurons incubated with FTLD-TDP Type A patient-brain derived TDP-43 filaments for 1 d and 3 d. Presynaptic active zone complex proteins are highlighted in red. B. Immunofluorescence Airyscan confocal microscopy images of mouse primary cortical neurons incubated with FTLD-TDP Type A patient-derived TDP-43 filaments for 3 d. Neurons were labelled using antibodies against microtubule-associated protein 2 (MAP2; grey); the presynaptic active zone complex protein bassoon (yellow); and PSD95 (cyan). TDP-43 filaments were labelled using the antibody against pTDP-43 (magenta). The arrow indicates an example of pTDP-43 signal colocalising with bassoon. Scale bars, 5 µm and 1 µm, as indicated. C. Percentage of MAP2-masked pTDP-43 signal colocalised with bassoon (left) and PSD95 (right), and vice versa, from (C) in three-dimensional space. Each data point represents one technical replicate. n= 2 biological replicates. Means +/- SD are shown.

Article Snippet: The grids were then blotted and incubated with anti-TDP-43 primary antibody (Proteintech, 10782-2-AP; 1:25) diluted in blocking buffer for 3 h at 21°C.

Techniques: Derivative Assay, Incubation, Immunofluorescence, Confocal Microscopy

Journal: bioRxiv

Article Title: Pathological TDP-43 filaments accumulate at synapses and cause synaptic dysfunction

doi: 10.64898/2026.01.27.701787

Figure Lengend Snippet: A. Immunoblot of synaptosome isolation fractions (s, supernatant; cp, cell pellet; cyt, cytosolic; syn, synaptosomes) from mouse primary cortical neurons incubated with (+ filaments) and without (- filaments) FTLD-TDP Type A patient brain-derived TDP-43 filaments for 3 d, probed with antibodies against postsynaptic density protein 95 (PSD95, top), pS409/410 TDP-43 (pTDP-43, middle) and TDP-43 (bottom). The arrows indicate full-length (FL) TDP-43 and C-terminal fragments (CTFs) of TDP-43. B. Denoised tomographic slices of synaptosomes from mouse primary cortical neurons incubated with FTLD-TDP Type A patient brain-derived TDP-43 filaments for 3 d. TDP-43 filaments (magenta arrows), example synaptic vesicles (cyan arrows) and the postsynaptic density (yellow arrow) are indicated. C and D. Denoised tomographic slices showing TDP-43 filaments (magenta arrows). making contacts (green arrows) with synaptic vesicles (cyan arrows) (C) and the presynaptic plasma membrane (yellow arrows) (D) . B-D. Scale bars, 50 nm.

Article Snippet: The grids were then blotted and incubated with anti-TDP-43 primary antibody (Proteintech, 10782-2-AP; 1:25) diluted in blocking buffer for 3 h at 21°C.

Techniques: Western Blot, Isolation, Incubation, Derivative Assay, Clinical Proteomics, Membrane

Journal: bioRxiv

Article Title: Pathological TDP-43 filaments accumulate at synapses and cause synaptic dysfunction

doi: 10.64898/2026.01.27.701787

Figure Lengend Snippet: A. Representative false-coloured images of mouse primary cortical neurons expressing SypHy prior to stimulation (Rest), during stimulation with a train of 300 action potentials delivered at 10 Hz (10 Hz), following stimulation (Recovery) and during challenge with ammonium chloride (NH 4 ) buffer to reveal the total synaptic vesicle pool. Arrows indicate examples of terminals that respond to stimulation. Scale bar, 2 μm. B. Time course of relative SypHy fluorescence intensity (ΔF/F0) in mouse primary cortical neurons incubated with FTLD-TDP Type A patient brain-derived TDP-43 filaments (+ filaments) or with aged-matched control brain extracts (+ control) for 3 d, normalised to challenge with NH 4 buffer (total synaptic vesicle pool). The bar indicates the period of stimulation. n = 4 biological replicates. Means +/- SEM are shown. C. Rate of activity-dependent relative SypHy fluorescence increase (ΔF/F0) over the first 6 seconds of stimulation (stimulation triggered at 30 s). n = 4 biological replicates. Means +/- SEM are shown. Linear regression analysis was performed (lines), **p=0.0055. D. Representative traces of excitatory postsynaptic currents (EPSCs) evoked by paired presynaptic stimuli (50 ms interval) from mouse primary hippocampal neurons incubated with (+ filaments) or without (- filaments) FTLD-TDP Type A patient brain-derived TDP-43 filaments or with aged-matched control brain extracts (+ control) for 7 d. E. Plot of the ratios of the second to the first EPSC (paired-pulse ratio; PPR) from mouse primary hippocampal neurons incubated with (+ filaments) or without (- filaments) FTLD-TDP Type A patient brain-derived TDP-43 filaments or aged-matched control brain extracts (+ control) for 3-4 d (d3/4) and 6-7 d (d6/7). Each data point represents one paired recording. Data points are colour coded by biological replicate. n = 3 biological replicates. Means +/- SD are shown. A two-way ANOVA with Tukey’s multiple comparison test was performed, *p<0.05, ****p<0.0001. F. Widefield fluorescence microscopy images of mouse primary cortical neurons expressing the fluorescent calcium indicator jRGECO1a (red) over 1200 ms. Images were taken at 100 ms intervals. Scale bar, 100 μm. G. The spontaneous firing frequency calculated from (F) for mouse primary cortical neurons incubated with (+ filaments) or without (- filaments) FTLD-TDP Type A patient brain-derived TDP-43 filaments or aged-matched control brain extracts (+ control) for 9 d (d9) and 14 d (d14). Each data point represents one time-lapse recording. Data points are colour coded by biological replicate. n = 5 biological replicates. Means +/- SD are shown. A two-way ANOVA with Tukey’s multiple comparison test was performed, **p<0.01, ****p<0.0001.

Article Snippet: The grids were then blotted and incubated with anti-TDP-43 primary antibody (Proteintech, 10782-2-AP; 1:25) diluted in blocking buffer for 3 h at 21°C.

Techniques: Expressing, Fluorescence, Incubation, Derivative Assay, Control, Activity Assay, Comparison, Microscopy

Journal: Pharmaceuticals

Article Title: Mazdutide Ameliorates Metabolic Dysfunction-Associated Steatotic Liver Disease by Modulating Endoplasmic Reticulum Stress, Improving Lipid Metabolism and Alleviating Inflammation

doi: 10.3390/ph19030371

Figure Lengend Snippet: Mazdutide suppresses ER stress via the PERK pathway in MASLD mouse liver tissue. ( A ) Representative immunohistochemical staining of GRP78 and CHOP in liver sections (scale bar = 50 μm, 200× magnification). ( B ) Representative Western blot images of key proteins in the PERK pathway from each experimental group. ( C ) Quantitative densitometric analysis of PERK and eIF2α phosphorylation (presented as the p-PERK/PERK and p-eIF2α/eIF2α ratios) and the protein levels of GRP78, ATF4, and CHOP. All values were normalized to β-actin. Data are presented as the mean ± SD ( n = 5). a p < 0.05 vs. the NCD group; b p < 0.05 vs. the HFD group. Animal groups are defined in .

Article Snippet: Primary antibodies for GRP78, ATF4, CHOP, and β-actin were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Immunohistochemical staining, Staining, Western Blot, Phospho-proteomics

Journal: Pharmaceuticals

Article Title: Mazdutide Ameliorates Metabolic Dysfunction-Associated Steatotic Liver Disease by Modulating Endoplasmic Reticulum Stress, Improving Lipid Metabolism and Alleviating Inflammation

doi: 10.3390/ph19030371

Figure Lengend Snippet: Mazdutide inhibits the PERK-eIF2α-ATF4-CHOP pathway in MASLD cells. ( A ) Representative Western blot images showing the protein levels of GRP78, p-PERK, PERK, p-eIF2α, eIF2α, ATF4, CHOP, and β-actin. ( B ) Quantitative densitometric analysis of the p-PERK/PERK and p-eIF2α/eIF2α ratios and the protein levels of GRP78, ATF4, and CHOP. The protein band intensities were normalized to β-actin. Data are presented as the mean ± SD ( n = 3). a p < 0.05 vs. the NCD group; b p < 0.05 vs. the HFD group. Cell groups are as defined in .

Article Snippet: Primary antibodies for GRP78, ATF4, CHOP, and β-actin were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Western Blot

Journal: Cancer Biology & Medicine

Article Title: Migration and invasion inhibitory protein inhibits M2 macrophage polarization to suppress colorectal cancer progression through the STING–NFκB2–IL10 axis

doi: 10.20892/j.issn.2095-3941.2025.0282

Figure Lengend Snippet: MIIP modulates the STING–NFκB2–IL10 signaling pathway in CRC cells. Immunofluorescence staining showing the effects of MIIP knockdown and MIIP overexpression on dsDNA levels (A) and the distribution and expression of STING (B) in SW480 and SW620 cells. (C) Western blots showing the effects of MIIP knockdown and overexpression on STING, TRAF3, p-p100, and p52 levels. (D) ELISA results showing the effects of MIIP knockdown and overexpression on IL10 levels in the cell culture medium. (E and F) Effects of MIIP knockdown and/or STING knockdown on p52 expression (E) and IL10 levels in the cell culture medium (F). All data are presented as the means ± SDs ( n = 3). * P < 0.05, ** P < 0.01, and *** P < 0.001; scale bar, 50 μm.

Article Snippet: The primary antibodies used included mouse anti-dsDNA (1:1000, ab270732; Abcam, Cambridge, MA, USA), rabbit anti-NFKB2/p52 (1:200, 15503-1-AP; Proteintech, Wuhan, Hubei, China), rabbit anti-TMEM173/STING (1:200, 19851-1-AP; Proteintech, Wuhan, Hubei, China), and rabbit anti-IL-10 antibodies (1:100, 20850-1-AP; Proteintech, Wuhan, Hubei, China).

Techniques: Immunofluorescence, Staining, Knockdown, Over Expression, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Cancer Biology & Medicine

Article Title: Migration and invasion inhibitory protein inhibits M2 macrophage polarization to suppress colorectal cancer progression through the STING–NFκB2–IL10 axis

doi: 10.20892/j.issn.2095-3941.2025.0282

Figure Lengend Snippet: MIIP inhibits tumor growth and metastasis and M2 macrophage infiltration in a syngeneic CRC mouse model through the STING–NFκB2–IL10 axis. (A) Tumors in situ (indicated by green arrows) and liver metastatic nodules (indicated by black arrows) were observed in a syngeneic CRC mouse model. Statistics for the size of tumors in situ (B) and the number of metastatic foci (C). (D) Statistical analysis of IL10 levels in mouse blood. (E) Representative images of HE and IHC staining for SATB2, MIIP, STING, IL10, p52, and CD163 (green arrows) in tumor tissues from the different groups. The bar graphs show the percentages of high/low STING (F), IL-10 (G), and p52 (H) expression in the different groups. (I) Comparison of the number of CD163-positive cells among the different groups. All data are presented as the means ± SDs ( n = 3). ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; scale bar, 50 μm.

Article Snippet: The primary antibodies used included mouse anti-dsDNA (1:1000, ab270732; Abcam, Cambridge, MA, USA), rabbit anti-NFKB2/p52 (1:200, 15503-1-AP; Proteintech, Wuhan, Hubei, China), rabbit anti-TMEM173/STING (1:200, 19851-1-AP; Proteintech, Wuhan, Hubei, China), and rabbit anti-IL-10 antibodies (1:100, 20850-1-AP; Proteintech, Wuhan, Hubei, China).

Techniques: In Situ, Immunohistochemistry, Expressing, Comparison

Journal: BMC Oral Health

Article Title: Synergistic proanthocyanidin-copper oxygen-generating microneedle enhances anti-inflammatory activity in dental pulp stem cells and macrophage efferocytosis

doi: 10.1186/s12903-026-07870-1

Figure Lengend Snippet: A : Fluorescence staining of ROS levels in macrophages cultured in the leachate of each hydrogel group; ( B ): Semi-quantitative analysis of ROS fluorescence staining; ( C , D , E) : Expression levels of IL-6, IL-1β, and TNF-α in macrophages cultured in the leachate of each hydrogel group; ( F) : Statistical analysis of macrophage efferocytosis index by immunofluorescence staining; G : Statistical analysis of macrophage efferocytosis index by flow cytometry; ( H ): Immunofluorescence staining to detect macrophage efferocytosis levels (blue: nuclei of macrophages; green: cell membrane of macrophages; red: apoptotic hDPSCs); ( I ): Flow cytometry detection of macrophage efferocytosis levels (FITC-A: F4/80-labeled macrophages; PE-A: CellTracker-labeled apoptotic cells)

Article Snippet: Macrophages (RAW264.7) were incubated for 24 h, and 1 mL of DMEM containing 200 ng/mL LPS was added to induce inflammation for 24 h, then 5 × 10 4 stained apoptotic cells and samples of different groups were added and co-cultured for 24 h. Then, it was fixed with 4% paraformaldehyde for 20 min, washed with PBS, blocked with goat serum for 20 min at room temperature, and incubated with F4/80 antibody (Bosterbio, China) overnight at 4°C.

Techniques: Fluorescence, Staining, Cell Culture, Expressing, Immunofluorescence, Flow Cytometry, Membrane, Labeling