Journal: Journal of Translational Medicine

Article Title: N-acetyltransferase 10 affects the proliferation of intrahepatic cholangiocarcinoma and M2-type polarization of macrophages by regulating C-C motif chemokine ligand 2

doi: 10.1186/s12967-024-05664-z

Figure Lengend Snippet: NAT10 polarizes macrophages toward the M2 type through CCL2 . (A) Macrophages were polarized by treating them with the supernatant of ICC cells for 24 h. (B) After co-culturing ICC cells and macrophages for 24 h, the macrophages underwent polarization. (C) Co-culturing ICC cells with macrophages for 24 h resulted in the polarization of macrophages towards the M2 phenotype. (D) Immunofluorescence showed that CD86 expression increased and CD163 expression decreased in NAT10-knockdown tumors ( n = 6). Scale bars: 50 μm. (E and F) Western blot and ELISA showed that NAT10 knockdown decreased CCL2 expression levels in ICC cells and cell supernatant. (G) CCL2-knockdown cell lines were constructed and verified at the protein level. (H) Flow cytometry confirmed that CCL2 knockdown reduced the polarization of macrophages toward M2. (I) Immunofluorescence showed that CD86 expression increased and CD163 expression decreased in CCL2-knockdown tumors ( n = 6). Scale bars: 50 μm. Data are representative of three or more independent experimental replicates. Data are displayed as the mean ± SD. P -values were determined by Student’s t-test and one-way ANOVA in panels. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ICC, intrahepatic cholangiocarcinoma; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation; ANOVA, analysis of variance

Article Snippet: The levels of CCL2 in the cell supernatants were determined using an enzyme-linked immunosorbent assay (ELISA) kit (E-EL-H6005, Elabscience).

Techniques: Immunofluorescence, Expressing, Knockdown, Western Blot, Enzyme-linked Immunosorbent Assay, Construct, Flow Cytometry, Standard Deviation

Journal: Journal of Nanobiotechnology

Article Title: Folate-modified biomimetic nanovesicles loaded with a PU.1 inhibitor alleviate atherosclerosis by suppressing inflammation

doi: 10.1186/s12951-025-03825-w

Figure Lengend Snippet: PU.1 is upregulated in macrophages of advanced atherosclerotic lesions and promotes inflammation through IL-1β/NF-κB signaling. ( A ) Boxplot showing expression levels of PU.1 in early versus advanced atherosclerotic plaques based on the GSE43292 dataset. ( B ) UMAP plot of single-cell RNA sequencing data depicting major immune and stromal cell populations in atherosclerotic lesions. ( C ) UMAP feature plot showing SPI1 (encoding PU.1) expression predominantly enriched in macrophages. ( D ) Representative immunofluorescence staining of PU.1 (green) and CD68 (red) in aortic root sections from chow diet– and high fat diet–fed mice. Nuclei were counterstained with DAPI (blue). Scale bar: 200 μm. ( E ) CUT&Tag analysis showing genome-wide binding of PU.1 in macrophages. Heatmap indicates PU.1 enrichment near transcription start sites (TSS). ( F ) Genomic distribution of PU.1 binding peaks identified by CUT&Tag. ( G – H ) Representative CUT&Tag tracks showing PU.1 binding at the promoters of pro-inflammatory cytokines. ( I ) Dual-luciferase reporter assay confirming the transcriptional activation of the IL-1β promoter by PU.1 overexpression (OE). ( J ) Western blot showing that PU.1 knockdown suppressed ox-LDL–induced IL-1β expression and NF-κB pathway activation (p-IκB and p-p65). ( K ) Western blot demonstrating that IL-1β knockdown reversed PU.1-induced NF-κB activation. ( L ) Western blot analysis showing that the PU.1 inhibitor DB1976 attenuated ox-LDL–induced IL-1β expression and NF-κB activation. ( M ) qRT-PCR analysis of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, MCP-1) in macrophages treated with ox-LDL with or without DB1976. One-way ANOVA with Tukey’s multiple comparison post hoc test was used for statistical analysis. Data are presented as the mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: ELISA kits for mouse IL-1β (E-EL-M0037), IL-6 (E-EL-M0044), TNF-α (E-EL-M3063), and MCP-1 (E-EL-M3001) were purchased from Elabscience (China).

Techniques: Expressing, RNA Sequencing, Immunofluorescence, Staining, Genome Wide, Binding Assay, Luciferase, Reporter Assay, Activation Assay, Over Expression, Western Blot, Knockdown, Quantitative RT-PCR, Comparison, Standard Deviation

Journal: Journal of Nanobiotechnology

Article Title: Folate-modified biomimetic nanovesicles loaded with a PU.1 inhibitor alleviate atherosclerosis by suppressing inflammation

doi: 10.1186/s12951-025-03825-w

Figure Lengend Snippet: The anti-atherosclerotic effects of the D-FNVs. ( A - D ) BMDMs were co-treated with ox-LDL (100 µg/mL) and various formulations (free DB1976, D-NVs, D-FNVs). The levels of IL-1β, IL-6, TNF-α, and MCP-1 in the supernatant were measured by ELISA. ( E , F ) Flow cytometry analysis and quantification of intracellular ROS levels in BMDM cells treated with ox-LDL (100 µg/mL) and various formulations (free DB1976, D-NVs, D-FNVs), respectively, at 2 mM DB1976 for 24 h. ( G , H ) Flow cytometry analysis and quantification of apoptosis rates in BMDMs treated with ox-LDL (100 µg/mL) and various formulations (free DB1976, D-NVs, D-FNVs) at 2 mM DB1976 for 24 h. One-way ANOVA with Tukey’s multiple comparison post hoc test was used for statistical analysis. Data are presented as the mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: ELISA kits for mouse IL-1β (E-EL-M0037), IL-6 (E-EL-M0044), TNF-α (E-EL-M3063), and MCP-1 (E-EL-M3001) were purchased from Elabscience (China).

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Comparison, Standard Deviation

Journal: Journal of Nanobiotechnology

Article Title: Folate-modified biomimetic nanovesicles loaded with a PU.1 inhibitor alleviate atherosclerosis by suppressing inflammation

doi: 10.1186/s12951-025-03825-w

Figure Lengend Snippet: D-FNVs ameliorated inflammation in an atherosclerotic mouse model. ( A , B ) Representative images of plaques within the aortic root subjected to immunofluorescent staining for the macrophage marker CD68. Scale bar: 100 μm. ( C – F ) Levels of IL-1β, IL-6, TNF-α, and MCP-1 in aortic tissues collected from atherosclerotic mice treated with various formulations (saline, DB1976, D-NVs, D-FNVs). ( G – J ) Levels of IL-1β, IL-6, TNF-α, and MCP-1 in blood serum collected from the same groups of atherosclerotic mice. The n values are all biological replicates. One-way ANOVA with Tukey’s multiple comparison post hoc test was used for statistical analysis. Data are presented as the mean ± standard deviation (SD). * p < 0.05, ** p < 0.01, and *** p < 0.001. **** p < 0.0001

Article Snippet: ELISA kits for mouse IL-1β (E-EL-M0037), IL-6 (E-EL-M0044), TNF-α (E-EL-M3063), and MCP-1 (E-EL-M3001) were purchased from Elabscience (China).

Techniques: Staining, Marker, Saline, Comparison, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Enhancement of Psoriasis Treatment by Phellodendri Chinensis Cortex Carbon Dots (PCC-CDs) Through Modulation of the HMGB1/TLR4/MAPK/NF-κB Pathway

doi: 10.2147/IJN.S578399

Figure Lengend Snippet: PCC-CDs spray regulate the HMGB1/TLR4/MAPK/NF-κB signaling pathway to ameliorate psoriasis-like dermatitis. ( A ) Serum levels of HMGB1, IFN-γ, and VEGF in mice. ( B ) Protein expression levels of HMGB1/TLR4/MAPK/NF-κB signaling pathway-related proteins in mouse skin tissues. Each group n = 3. Data were expressed as means ± standard deviation (SD). # P < 0.05, ## P < 0.01, ### P < 0.001 vs Control group; * P < 0.05, ** P < 0.01, *** P < 0.001vs Model group.

Article Snippet: The following antibodies were used: PCNA antibody (Cat. 10205-2-AP, Proteintech, Wuhan, China), Ki67 Polyclonal antibody (Cat. 28074-1-AP, Proteintech, Wuhan, China), HMGB1 antibody (Cat. 10829-1-AP, Proteintech, Wuhan, China), TLR4 antibody (Cat. 30400-1-AP, Proteintech, Wuhan, China), phospho-p38 MAPK (Thr180/Tyr182) Polyclonal antibody (Cat. 28796-1-AP, Proteintech, Wuhan, China), p38 MAPK Polyclonal antibody (Cat. 14064-1-AP, Proteintech, Wuhan, China), phospho-NF-κB p65 (Ser468) Recombinant antibody (Cat. 82335-1-AP, Proteintech, Wuhan, China), NF-κB p65 Polyclonal antibody (Cat. 10745-1-AP, Proteintech, Wuhan, China), GAPDH antibody (Cat. 60004-1-Ig, Proteintech, Wuhan, China), HRP-conjugated Goat Anti-Rabbit lgG (Cat. SA00001-2, Proteintech, Wuhan, China), HRP-conjugated Goat Anti-Mouse lgG (Cat. SA00001-1, Proteintech, Wuhan, China), Pentobarbital sodium salt (Cat. P3761, Sigma-Aldrich, St. Louis, MO,USA).

Techniques: Expressing, Standard Deviation, Control

Journal: International Journal of Nanomedicine

Article Title: Enhancement of Psoriasis Treatment by Phellodendri Chinensis Cortex Carbon Dots (PCC-CDs) Through Modulation of the HMGB1/TLR4/MAPK/NF-κB Pathway

doi: 10.2147/IJN.S578399

Figure Lengend Snippet: PCC-CDs alleviate M1 macrophage polarisation by modulating the HMGB1/TLR4/NF-κB signalling pathway. ( A ) Cytokine levels in cell supernatants, including HMGB1, TNF-α, IL-1β, IL-6 and IL-10. ( B ) The effects of PCC-CD intervention on HMGB1/TLR4/NF-κB protein expression in M1 macrophages (n = 3 per group). Data are expressed as mean ± standard deviation (SD). # P < 0.05, ## P < 0.01 compared with the control group; *P < 0.05, **P < 0.01 compared with the model group.

Article Snippet: The following antibodies were used: PCNA antibody (Cat. 10205-2-AP, Proteintech, Wuhan, China), Ki67 Polyclonal antibody (Cat. 28074-1-AP, Proteintech, Wuhan, China), HMGB1 antibody (Cat. 10829-1-AP, Proteintech, Wuhan, China), TLR4 antibody (Cat. 30400-1-AP, Proteintech, Wuhan, China), phospho-p38 MAPK (Thr180/Tyr182) Polyclonal antibody (Cat. 28796-1-AP, Proteintech, Wuhan, China), p38 MAPK Polyclonal antibody (Cat. 14064-1-AP, Proteintech, Wuhan, China), phospho-NF-κB p65 (Ser468) Recombinant antibody (Cat. 82335-1-AP, Proteintech, Wuhan, China), NF-κB p65 Polyclonal antibody (Cat. 10745-1-AP, Proteintech, Wuhan, China), GAPDH antibody (Cat. 60004-1-Ig, Proteintech, Wuhan, China), HRP-conjugated Goat Anti-Rabbit lgG (Cat. SA00001-2, Proteintech, Wuhan, China), HRP-conjugated Goat Anti-Mouse lgG (Cat. SA00001-1, Proteintech, Wuhan, China), Pentobarbital sodium salt (Cat. P3761, Sigma-Aldrich, St. Louis, MO,USA).

Techniques: Expressing, Standard Deviation, Control

Journal: International Journal of Nanomedicine

Article Title: Enhancement of Psoriasis Treatment by Phellodendri Chinensis Cortex Carbon Dots (PCC-CDs) Through Modulation of the HMGB1/TLR4/MAPK/NF-κB Pathway

doi: 10.2147/IJN.S578399

Figure Lengend Snippet: Schematic illustration of the therapeutic mechanism hypothesis of PCC-CDs spray in alleviating psoriatic inflammation via modulation of the HMGB1/TLR4/MAPK/NF-κB signaling pathway.

Article Snippet: The following antibodies were used: PCNA antibody (Cat. 10205-2-AP, Proteintech, Wuhan, China), Ki67 Polyclonal antibody (Cat. 28074-1-AP, Proteintech, Wuhan, China), HMGB1 antibody (Cat. 10829-1-AP, Proteintech, Wuhan, China), TLR4 antibody (Cat. 30400-1-AP, Proteintech, Wuhan, China), phospho-p38 MAPK (Thr180/Tyr182) Polyclonal antibody (Cat. 28796-1-AP, Proteintech, Wuhan, China), p38 MAPK Polyclonal antibody (Cat. 14064-1-AP, Proteintech, Wuhan, China), phospho-NF-κB p65 (Ser468) Recombinant antibody (Cat. 82335-1-AP, Proteintech, Wuhan, China), NF-κB p65 Polyclonal antibody (Cat. 10745-1-AP, Proteintech, Wuhan, China), GAPDH antibody (Cat. 60004-1-Ig, Proteintech, Wuhan, China), HRP-conjugated Goat Anti-Rabbit lgG (Cat. SA00001-2, Proteintech, Wuhan, China), HRP-conjugated Goat Anti-Mouse lgG (Cat. SA00001-1, Proteintech, Wuhan, China), Pentobarbital sodium salt (Cat. P3761, Sigma-Aldrich, St. Louis, MO,USA).

Techniques:

Journal: Frontiers in Medicine

Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

doi: 10.3389/fmed.2026.1719787

Figure Lengend Snippet: Chronic ethanol ingestion altered the oscillation patterns of key circadian signaling molecules in mouse lungs. Graphs summarize expression profiles of key circadian regulators in mouse lungs at 4 h intervals over 24 h for ethanol-fed (EtOH, red) and control-fed (CON, blue) groups. Panels show expressions of (A) Bmal1 , (B) Clock , (C) Rorα , and (D) Rev-erbα analyzed by qPCR. N = 6–9 lungs per group. The circadian oscillation patterns of the four genes from ethanol-fed mouse lungs significantly changed compared to lungs from control-fed mice. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control-fed (CON) group at the same time point.

Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

Techniques: Expressing, Control

Journal: Frontiers in Medicine

Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

doi: 10.3389/fmed.2026.1719787

Figure Lengend Snippet: Chronic ethanol ingestion disrupts temporal protein expression of key circadian signaling molecules in the mouse lungs. Protein levels from mouse lungs collected at ZT0 and ZT12 were analyzed for (A) BMAL1 (75 kD), (B) CLOCK (100 kD), (C) RORα (60 kD), and (D) REV-ERBα (75 kD) and normalized with GAPDH (36 kD) levels in the same blot by western analysis. All data reported as fold-change compared to control-fed at ZT0 (CON) group. Representative western immunoblots are shown above the corresponding graphs. N = 6 per group. Data are presented as mean ± SE. # indicates change with p < 0.005 compared with ZT0 CON group and * indicates changes with p < 0.05 compared with ZT12 CON group.

Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

Techniques: Expressing, Western Blot, Control

Journal: Frontiers in Medicine

Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

doi: 10.3389/fmed.2026.1719787

Figure Lengend Snippet: Ethanol exposure in vitro inhibited key molecules in the circadian signaling pathway. Gene expression levels from synchronized and unsynchronized murine primary lung fibroblasts (mPLFs) ± ethanol (EtOH, 60 mM; 24 h) were analyzed for: (A,B) Bmal1 , (C,D) Clock , (E,F) Rorα , and (G,H) Rev-erbα mRNA expression. Ethanol exposure significantly inhibited BMAL1, RORα, and Rev-erbα gene expressions while upregulating Clock gene expression. N = 6–10 per group. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control (CON) group.

Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

Techniques: In Vitro, Gene Expression, Expressing, Control

Journal: Frontiers in Medicine

Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

doi: 10.3389/fmed.2026.1719787

Figure Lengend Snippet: Ethanol treatment inhibited core circadian signaling molecules in murine lung fibroblasts. Protein levels from synchronized and unsynchronized murine primary lung (mPLFs) ± ethanol (EtOH, 60 mM; 72 h) were analyzed for: (A,B) BMAL1 (75 kD), (C,D) CLOCK (100 kD), (E,F) RORα (60 kD), and (G,H) REV-ERBα (75 kD) were normalized to GAPDH (36 kD) levels from the same blot and reported as fold-change compared to the untreated (CON) group. Ethanol treatment significantly decreased the BAML1 and RORα protein levels in both synchronized and unsynchronized conditions, decreased REV-ERBα in the unsynchronized condition while it did not change the CLOCK protein expression in either condition. Representative western immunoblots are shown above the graphs. N = 8–15 per group. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control (CON) group.

Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

Techniques: Expressing, Western Blot, Control