Journal: International Journal of Molecular Medicine

Article Title: Selective autophagic degradation of glycolytic activator PFKFB3 contributes to maintaining intestinal epithelial barrier in inflammatory bowel disease

doi: 10.3892/ijmm.2025.5714

Figure Lengend Snippet: Impaired autophagy and upregulated glycolysis levels in IECs were observed under inflammatory conditions. (A) Establishment of acute colitis mouse model and chronic colitis mouse model. (B) The protein levels of P62 and LC3b in colonic epithelial cells of acute DSS-induced mice (n=3) and chronic DSS-induced mice (n=3) determined by western blot analysis. (C) mRNA levels of PFKP, LDHA, HK2, PKM2, PFKFB3 in DSS-induced mice epithelial cells. (D) Autophagic flux was measured by transfecting cells with mRFP-GFP-LC3 dual-fluorescence adenovirus (Ad-LC3), allowing differentiation between autophagosomes (mRFP+/GFP+ fluorescence, appearing as yellow puncta) and autolysosomes (mRFP+/GFP-fluorescence, appearing as red puncta). TNF-α (200 ng/ml) induced epithelial cells for 48 h. Representative immunofluorescence images were shown. (Original magnification, ×20). Quantification of LC3 puncta number of representative cells. (E) Relative glucose content and lactate (Lac) production in control vs. TNF-α induced epithelial cells. (F) 2-NBDG and (G) BCECF fluorescent counter-staining in TNF-α induced epithelial cells vs. control. Representative immunofluorescence images were shown. (Original magnification, ×20). Quantitative fluorescence intensity of 2-NBDG and BCECF (n=3). Data were presented as the mean ± SD of n=3 per group. Statistical analysis was performed using unpaired t-tests. **** P<0.0001, *** P<0.001, ** P<0.01, * P<0.05, ns, non-significant vs. control group. IECs, intestinal epithelial cells; DSS, dextran sulfate sodium salt; 2-NBDG, 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]-D-glucose; BCECF, 2',7'-bis(carboxyethyl)-5-carboxyfluorescein.

Article Snippet: TNF-α (HY-P70426A) was purchased from MedChemExpress.

Techniques: Western Blot, Fluorescence, Immunofluorescence, Control, Staining

Journal: International Journal of Molecular Medicine

Article Title: Selective autophagic degradation of glycolytic activator PFKFB3 contributes to maintaining intestinal epithelial barrier in inflammatory bowel disease

doi: 10.3892/ijmm.2025.5714

Figure Lengend Snippet: PFKFB3 was upregulated in IBD. (A) Analysis of PFKFB3 mRNA expression in normal and Crohn's disease tissues within public datasets ( GSE119600 , GSE126124 , GSE112057 ). (B) mRNA levels of PFKPFB3 in control (n=10) vs. tissue of patients with Crohn's disease (n=10), control (n=26) vs. blood of patients with Crohn's disease (active stage n=12; remission stage n=7). (C) Immunohistochemical staining of PFKFB3 in tissue of patients with Crohn's disease and semiquantitative analysis (n=3). (D) Immunofluorescence images of PFKFB3 in colonic epithelial tissue of patients with Crohn's disease and quantification of the fluorescence intensity of PFKFB3 (n=3). (E) Protein levels of PFKFB3 in control vs. colonic epithelium of DSS-induced mice (acute n=3, chronic n=3). (F) Immunofluorescence images of PFKFB3 in colonic epithelium of DSS-induced mice and quantification of the fluorescence intensity of PFKFB3 (acute n=3, chronic n=3). (G) Representative hematoxylin and eosin staining of the colon. DSS-induced colitis mice were treated with intraperitoneal injection of PFK-15 every three days. (H) Immunohistochemical staining of claudin-5, claudin-8, claudin-2 and occludin in colon tissue. (I) mRNA levels of IL-6, TNF-α and IL-1β. The P values for all figures except B, F, and H were calculated using unpaired t-tests, while one-way ANOVA followed by Tukey's multiple comparisons test was used for B, F and H. The values were presented as the means ± SEM, * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. PFKFB3, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3; IBD, inflammatory bowel disease; DSS, dextran sulfate sodium salt; PFK-1, phosphofructokinase-1; CD, Crohn's disease; WT, Wild-type.

Article Snippet: TNF-α (HY-P70426A) was purchased from MedChemExpress.

Techniques: Expressing, Control, Immunohistochemical staining, Staining, Immunofluorescence, Fluorescence, Injection

Journal: Chinese Medicine

Article Title: Single-cell RNA sequencing reveals the therapeutic mechanism of Calvatia lilacina in promoting wound healing of anal fistula

doi: 10.1186/s13020-025-01293-w

Figure Lengend Snippet: The impact of CLS on inflammation, angiogenesis, and collagen at different time points. A – C ELISA detection of TNF-α, VEGF, and Collagen I expression levels in granulation tissue; D Immunohistochemical staining of CD14 and CD11b; E , F Immunohistochemical quantitative analysis. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and ns: not significant ( P > 0.05) vs.the control group; n = 10

Article Snippet: TNF-α, COL1A1, VEGF Elisa Kit (EK182, EK183, EK1C01, Multi Sciences, China); WASF3 Antibody (67620-1-lg, Proteintech, China); α-SMA (19245s, CST, USA); CD14 Antibody (ab183322, abcam, Britain); Cluster of Differentiation 11b (CD11b) Antibody (ab133357, abcam, Britain); IL-6 flow Antibody (562050, BD Pharmingen, USA); JAK2 (WL02188, Wanleibio, China); p-JAK2 (WL02997, Wanleibio, China); STAT3 (WL03207, Wanleibio, China); p-STAT3 (WL06214, Wanleibio, China);

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemical staining, Staining, Control

Journal: Chinese Medicine

Article Title: Single-cell RNA sequencing reveals the therapeutic mechanism of Calvatia lilacina in promoting wound healing of anal fistula

doi: 10.1186/s13020-025-01293-w

Figure Lengend Snippet: Flow cytometry and SEM detection of adhesion and effect of CLS on macrophages. A Flow cytometry was used to determine the ratio of IL-6 positive cells in macrophage after extraction. B Quantitative analysis the rate of IL-6 positive macrophage. C , D Elisa detection of IL-6 and CXCL-8 in cell supernatant. E , F Intensity of intercellular communication between control group and treatment group. G Analysis of IL-6 + macrophage and fibroblast receptor-ligand binding. H Analysis of cell population interaction intensity of IL-6, CXCL-8 signaling pathway. * P < 0.05, ** P < 0.01, *** P < 0.001, and ns: not significant ( P > 0.05) vs.the control group; n = 3

Article Snippet: TNF-α, COL1A1, VEGF Elisa Kit (EK182, EK183, EK1C01, Multi Sciences, China); WASF3 Antibody (67620-1-lg, Proteintech, China); α-SMA (19245s, CST, USA); CD14 Antibody (ab183322, abcam, Britain); Cluster of Differentiation 11b (CD11b) Antibody (ab133357, abcam, Britain); IL-6 flow Antibody (562050, BD Pharmingen, USA); JAK2 (WL02188, Wanleibio, China); p-JAK2 (WL02997, Wanleibio, China); STAT3 (WL03207, Wanleibio, China); p-STAT3 (WL06214, Wanleibio, China);

Techniques: Flow Cytometry, Extraction, Enzyme-linked Immunosorbent Assay, Control, Ligand Binding Assay

Journal: International Journal of Molecular Medicine

Article Title: TWEAK modulates the characteristics of periodontal ligament stem cells via the Fn14/NF-κB pathway

doi: 10.3892/ijmm.2025.5679

Figure Lengend Snippet: Effects of TWEAK on the proliferation and apoptosis of PDLSCs. (A) Line chart depicting the effects of various concentrations of TWEAK (0, 1, 5, 20, 50 and 100 ng/ml) on PDLSC proliferation over 5 days, as assessed using the CCK-8 assay. (B) Statistical analysis of the CCK-8 data from day 5 of TWEAK stimulation shown in (A). (C) Statistical analysis of the average fluorescence intensity of TUNEL (green) following treatment with varying concentrations of TWEAK (0, 1, 5, 20, 50 and 100 ng/ml). (D) TUNEL assay detecting the effects of various concentrations (0, 1, 5, 20, 50 and 100 ng/ml) of TWEAK on PDLSC apoptosis. Scale bar, 200 μ m. Statistical analysis was performed using a one-way ANOVA. * P<0.05; **** P<0.0001. Data are presented as the mean ± SD (n=5 or 6). CCK-8, Cell Counting Kit-8; OD450, optical density at 450 nm; PDLSC, periodontal ligament stem cell; TWEAK, tumor necrosis factor-like weak inducer of apoptosis.

Article Snippet: Recombinant human cynomolgus TWEAK/TNF superfamily member 12 (TNFSF12) protein (mFc tag) (cat. no. 90094-C04H; Sino Biological, Inc.) was dissolved in sterile water to obtain a stock solution concentration of 50 μ g/ml.

Techniques: CCK-8 Assay, Fluorescence, TUNEL Assay, Cell Counting

Journal: International Journal of Molecular Medicine

Article Title: TWEAK modulates the characteristics of periodontal ligament stem cells via the Fn14/NF-κB pathway

doi: 10.3892/ijmm.2025.5679

Figure Lengend Snippet: Effects of TWEAK on the migration and osteogenic differentiation of PDLSCs. (A) Effects of various concentrations of TWEAK (0, 1, 5, 20, 50 and 100 ng/ml) on the number of migrating PDLSCs, and (B) quantitative analysis of the number of migrating cells (n=6). Scale bar, 400 μ m. (C) Effects of various concentrations of TWEAK on PDLSC migration toward scratch wounds over 24 h, and (D) quantitative analysis of the percentage of wound area reduction (n=12). Scale bar, 1 mm. (E) Effects of various concentrations of TWEAK on ALP staining in PDLSCs, and (F) quantitative analysis of grayscale values from ALP staining (n=6). Scale bar, 200 μ m. (G) Alizarin Red staining revealed the effects of various concentrations of TWEAK on PDLSC mineralization, and (H) quantitative analysis of grayscale values from Alizarin Red staining (n=6). Scale bar, 200 μ m. (I) Reverse transcription-quantitative PCR was used to assess the mRNA expression levels of RUNX2 , SP7 , ALP and OPG in PDLSCs after TWEAK stimulation (n=4), with β-actin serving as the internal control. (J) Western blot analysis of RUNX2, SP7, ALP and OPG protein expression in PDLSCs after TWEAK induction, and (K) semi-quantitative analysis of the gel band intensity, using β-actin as the internal control. Statistical analysis was performed using a one-way ANOVA. * P<0.05; ** P<0.01; *** P<0.001; **** P<0.0001. Data are presented as the mean ± SD. ALP, alkaline phosphatase; IOD, integral optical density; ns, not significant; OPG, osteoprotegerin; PDLSC, periodontal ligament stem cell; RUNX2, runt-related transcription factor 2; SP7, Sp7 transcription factor; TWEAK, tumor necrosis factor-like weak inducer of apoptosis.

Article Snippet: Recombinant human cynomolgus TWEAK/TNF superfamily member 12 (TNFSF12) protein (mFc tag) (cat. no. 90094-C04H; Sino Biological, Inc.) was dissolved in sterile water to obtain a stock solution concentration of 50 μ g/ml.

Techniques: Migration, Staining, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Control, Western Blot

Journal: International Journal of Molecular Medicine

Article Title: TWEAK modulates the characteristics of periodontal ligament stem cells via the Fn14/NF-κB pathway

doi: 10.3892/ijmm.2025.5679

Figure Lengend Snippet: Transcriptome analysis of the effect of TWEAK treatment on PDLSCs. (A) Venn diagram illustrating differentially expressed genes in PDLSCs following TWEAK treatment. (B) Multipoint differential scatter plot showing differentially expressed genes in PDLSCs following TWEAK treatment. (C) GO enrichment analysis comparing PDLSCs and PDLSCs treated with 50 ng/ml TWEAK. (D) KEGG enrichment analysis comparing PDLSCs and PDLSCs treated with 50 ng/ml TWEAK. (E) GO enrichment analysis comparing PDLSCs and PDLSCs treated with 100 ng/ml TWEAK. (F) KEGG enrichment analysis comparing PDLSCs and PDLSCs treated with 100 ng/ml TWEAK. (G) Western blot analysis was conducted to detect the levels of Fn14, NF-κB, P-NF-κB and NLRP3 in PDLSCs stimulated with 50 and 100 ng/ml TWEAK. (H) Statistical analysis of protein band intensities from (G) (n=3). (I) Western blot analysis of the levels of Fn14, NF-κB, P-NF-κB and NLRP3 in PDLSCs after Fn14 was silenced using an shRNA. (J) Statistical analysis of protein band intensities from (I) (n=3). Statistical analysis was performed using (G and H) one-way ANOVA or (I and J) a two-tailed Student's t-test. * P<0.05; ** P<0.01; *** P<0.001. Data are presented as the mean ± SD. FC, fold change; Fn14, fibroblast growth factor-inducible 14; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; NLRP3, NOD-like receptor thermal protein domain-associated protein 3; ns, not significant; P-, phosphorylated; PDLSC, periodontal ligament stem cell; shRNA/sh, short hairpin RNA; TWEAK, tumor necrosis factor-like weak inducer of apoptosis.

Article Snippet: Recombinant human cynomolgus TWEAK/TNF superfamily member 12 (TNFSF12) protein (mFc tag) (cat. no. 90094-C04H; Sino Biological, Inc.) was dissolved in sterile water to obtain a stock solution concentration of 50 μ g/ml.

Techniques: Western Blot, shRNA, Two Tailed Test

Journal: International Journal of Molecular Medicine

Article Title: TWEAK modulates the characteristics of periodontal ligament stem cells via the Fn14/NF-κB pathway

doi: 10.3892/ijmm.2025.5679

Figure Lengend Snippet: Inhibition of Fn14 and NF-κB effectively blocks TWEAK-induced alterations in PDLSC characteristics. (A) Western blot analysis was conducted to assess the levels of Fn14, NF-κB, P-NF-κB and NLRP3 in PDLSCs. (B) Statistical analysis of the intensities of the protein bands shown in (A) (n=3). (C) A CCK-8 assay was performed to generate the proliferation curve of PDLSCs. (D) Statistical analysis of the OD450 values of cells from each group on day 5 of the CCK-8 assay, as presented in (C) (n=6). (G) Results of ALP staining and (E) the corresponding statistical analysis of PDLSCs after osteogenic induction (n=6). Scale bar, 400 μ m. (H) Results of Alizarin Red staining and (F) the corresponding statistical analysis of PDLSCs following osteogenic induction (n=6). Scale bar, 400 μ m. (I) Reverse transcription-quantitative PCR was used to assess the mRNA expression levels of RUNX2 , SP7 , ALP and OPG in PDLSCs, with β-actin serving as an internal reference (n=4). (J) Western blot analysis was performed to detect the protein expression levels of RUNX2, SP7, ALP and OPG in PDLSCs, and (K) the grayscale values of the gel images were semi-quantitatively analyzed, with β-actin used as an internal reference (n=3). Statistical analysis was performed using a one-way ANOVA. * P<0.05; ** P<0.01; *** P<0.001; **** P<0.0001. Data are presented as the mean ± SD. ALP, alkaline phosphatase; CCK-8, Cell Counting Kit-8; Fn14, fibroblast growth factor-inducible 14; IOD, integral optical density; NLRP3, NOD-like receptor thermal protein domain-associated protein 3; ns, not significant; OD450, optical density at 450 nm; OPG, osteoprotegerin; P-, phosphorylated; PDLSC, periodontal ligament stem cell; RUNX2, runt-related transcription factor 2; sh, short hairpin RNA; SP7, Sp7 transcription factor; TWEAK, tumor necrosis factor-like weak inducer of apoptosis.

Article Snippet: Recombinant human cynomolgus TWEAK/TNF superfamily member 12 (TNFSF12) protein (mFc tag) (cat. no. 90094-C04H; Sino Biological, Inc.) was dissolved in sterile water to obtain a stock solution concentration of 50 μ g/ml.

Techniques: Inhibition, Western Blot, CCK-8 Assay, Staining, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Cell Counting, shRNA

Journal: International Journal of Molecular Medicine

Article Title: TWEAK modulates the characteristics of periodontal ligament stem cells via the Fn14/NF-κB pathway

doi: 10.3892/ijmm.2025.5679

Figure Lengend Snippet: Inhibition of Fn14 and NF-κB effectively blocks TWEAK-induced alterations in the microenvironmental regulatory potential of PDLSCs. (A) Expression levels of OPG (green) and RANKL (red) in PDLSCs were detected by immunofluorescence staining, followed by quantitative analysis of the MOD values for (B) RANKL and (C) OPG, and (D) the MOD ratio of RANKL/OPG (n=5). Scale bar, 200 μ m. (E) Expression levels of CD68 and CD163 in RAW264.7 macrophages were detected by immunofluorescence staining, followed by quantitative analysis of the MOD values for (F) CD68 and (G) CD163 (n=5). Scale bar, 200 μ m. Statistical analysis was performed using a one-way ANOVA. ** P<0.01; *** P<0.001; **** P<0.0001. Data are presented as the mean ± SD. Fn14, fibroblast growth factor-inducible 14; MOD, mean optical density; ns, not significant; OPGa, osteoprotegerin; PDLSC, periodontal ligament stem cell; RANKL, receptor activator of nuclear factor-κB ligand; sh, short hairpin RNA; TWEAK, tumor necrosis factor-like weak inducer of apoptosis.

Article Snippet: Recombinant human cynomolgus TWEAK/TNF superfamily member 12 (TNFSF12) protein (mFc tag) (cat. no. 90094-C04H; Sino Biological, Inc.) was dissolved in sterile water to obtain a stock solution concentration of 50 μ g/ml.

Techniques: Inhibition, Expressing, Immunofluorescence, Staining, shRNA

Journal: International Journal of Molecular Medicine

Article Title: TWEAK modulates the characteristics of periodontal ligament stem cells via the Fn14/NF-κB pathway

doi: 10.3892/ijmm.2025.5679

Figure Lengend Snippet: Inhibition of the TWEAK/Fn14/NF-κB/NLRP3 pathway enhances the functional properties of iPDLSCs. (A) Expression profile of surface markers in iPDLSCs quantified using flow cytometry. (B) Levels of TWEAK, Fn14, NF-κB, P-NF-κB and NLRP3 in PDLSCs, iPDLSCs, and iPDLSCs after the downregulation of Fn14, NF-κB and NLRP3, and (C) statistical analysis of the band density values (n=3). (D) Apoptosis levels in PDLSCs, iPDLSCs, and iPDLSCs after downregulation of Fn14, NF-κB and NLRP3 were detected using the TUNEL assay, and (E) statistical analysis of the average fluorescence intensity of TUNEL was performed (n=5). Scale bar, 200 μ m. (F) A Cell Counting Kit-8 assay was used to assess the proliferative potential of PDLSCs, iPDLSCs, and iPDLSCs after the downregulation of Fn14, NF-κB and NLRP3, and (G) statistical analysis of the OD450 values on day 5 of the experiment was performed (n=6). (H) Transwell migration assay evaluating the migratory potential of PDLSCs, iPDLSCs, and iPDLSCs after Fn14, NF-κB or NLRP3 downregulation, with (I) quantification of the number of migrated cells (n=6). Scale bar, 400 μ m. (J) Wound healing assay evaluating the migratory potential of PDLSCs, iPDLSCs, and iPDLSCs after Fn14, NF-κB or NLRP3 downregulation, with (K) quantification of the percentage of wound closure (%) (n=16). Scale bar, 1 mm. (L) ALP staining was used to evaluate the mineralization potential of PDLSCs, iPDLSCs, and iPDLSCs after downregulation of Fn14, NF-κB or NLRP3, with (M) quantification of the integral optical density of the ALP-stained images (n=6). Scale bar, 400 μ m. (N) Alizarin Red staining was used to evaluate the mineralization potential of PDLSCs, iPDLSCs, and iPDLSCs after downregulation of Fn14, NF-κB or NLRP3, with (O) quantification of the integral optical density of the Alizarin Red-stained images (n=6). Scale bar, 400 μ m. (P) Reverse transcription-quantitative PCR was used to evaluate the mRNA expression levels of RUNX2 , SP7 , ALP and OPG in PDLSCs, iPDLSCs, and iPDLSCs after the downregulation of Fn14, NF-κB and NLRP3 (n=4). (Q) Western blotting was used to detect the expression levels of RUNX2, SP7, ALP and OPG in PDLSCs, iPDLSCs, and iPDLSCs after the downregulation of Fn14, NF-κB and NLRP3, and (R) statistical analysis of the band density values was performed (n=3). Statistical analysis was performed using a one-way ANOVA. * P<0.05; ** P<0.01; *** P<0.001; **** P<0.0001. Data are presented as the mean ± SD. ALP, alkaline phosphatase; Fn14, fibroblast growth factor-inducible 14; IOD, integral optical density; iPDLSC, inflammatory PDLSC; NLRP3, NOD-like receptor thermal protein domain-associated protein 3; ns, not significant; OD450, optical density at 450 nm; OPG, osteoprotegerin; P-, phosphorylated; PDLSC, periodontal ligament stem cell; RUNX2, runt-related transcription factor 2; sh, short hairpin RNA; SP7, Sp7 transcription factor; TWEAK, tumor necrosis factor-like weak inducer of apoptosis.

Article Snippet: Recombinant human cynomolgus TWEAK/TNF superfamily member 12 (TNFSF12) protein (mFc tag) (cat. no. 90094-C04H; Sino Biological, Inc.) was dissolved in sterile water to obtain a stock solution concentration of 50 μ g/ml.

Techniques: Inhibition, Functional Assay, Expressing, Flow Cytometry, TUNEL Assay, Fluorescence, Cell Counting, Transwell Migration Assay, Wound Healing Assay, Staining, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, shRNA

Journal: International Journal of Molecular Medicine

Article Title: TWEAK modulates the characteristics of periodontal ligament stem cells via the Fn14/NF-κB pathway

doi: 10.3892/ijmm.2025.5679

Figure Lengend Snippet: Effects of TWEAK and TWEAK-Fn14-IN-1 on the progression of rat periodontitis. (A) Micro-CT images of the rat maxilla, with the distance between the two red short lines representing the CEJ-ABC distance on the buccal side, and with statistical analysis of the (B) distance of CEJ-ABC (n=6), and (C) BV/TV at the root bifurcation of the maxillary second molar (n=6). Scale bar, 1 mm. Images of (D) H&E and (E) Masson's trichrome staining of rat periodontal tissues. Scale bar, 1 mm (top) or 100 μ m (bottom). (F) Representative images of TRAP/alkaline phosphatase double staining in the periodontal tissue of the rat maxillary second molar, with (G) quantification and statistical analysis of osteoclast numbers (TRAP-positive, multinucleated cells located in the bone resorption lacunae) at the mesial root (n=6). Red triangles indicate osteoclasts. Scale bar, 50 μ m. (H) Immunofluorescence staining of CD163 (green) and CD68 (red) in periodontal tissues, with (J) quantification of the mean fluorescence intensity of CD163 and (K) quantification of the mean fluorescence intensity of CD68 (n=6). Scale bar, 100 μ m. (I) Immunofluorescence staining of RUNX2 (green) and Periostin (red) in periodontal tissues, with (L) quantification of the mean fluorescence intensity of RUNX2 and (M) quantification of the mean fluorescence intensity of Periostin (n=6). Scale bar, 100 μ m. Blank represents the unmodeled group, PBS refers to the control group where PBS was used instead of TWEAK or TWEAK-Fn14-IN-1 during modeling, and TWEAK and TWEAK-Fn14-IN-1 represent experimental groups where the recombinant TWEAK protein or TWEAK-Fn14-IN-1 inhibitor was applied, respectively. Statistical analysis was performed using a one-way ANOVA. * P<0.05; ** P<0.01; **** P<0.0001. Data are presented as the mean ± SD. ABC, alveolar bone crest; BV/TV, bone volume to total volume; CEJ, cementoenamel junction; Fn14, fibroblast growth factor-inducible 14; MOD, mean optical density; ns, not significant; RUNX2, runt-related transcription factor 2; TRAP, tartrate-resistant acid phosphatase; TWEAK, tumor necrosis factor-like weak inducer of apoptosis.

Article Snippet: Recombinant human cynomolgus TWEAK/TNF superfamily member 12 (TNFSF12) protein (mFc tag) (cat. no. 90094-C04H; Sino Biological, Inc.) was dissolved in sterile water to obtain a stock solution concentration of 50 μ g/ml.

Techniques: Micro-CT, Staining, Double Staining, Immunofluorescence, Fluorescence, Control, Recombinant

Journal: Microbiology Spectrum

Article Title: Cathepsin S contributes to influenza-induced lung injury by driving inflammation, promoting apoptosis, and disrupting epithelial barrier integrity

doi: 10.1128/spectrum.01128-25

Figure Lengend Snippet: CTSS does not affect PR8 virus replication but reduces cytokine expression in A549 cells. A549 cells were transfected with two different CTSS-siRNAs (siCTSS #1 and siCTSS #2) or a scramble siRNA (siSCR) and then infected with 0.25 MOI of PR8 virus. ( A ) Left panel: representative western Blot analysis of CTSS and viral proteins (HA and NP) expression at 24 hpi, Right panel: intensity scanning of the bands from the left, quantified with data from three independent repeats. ( B ) Viral M gene expression at 24 hpi was quantified by RT-qPCR. ( C ) Viral titers at specified timepoints were measured using the TCID 50 assay. ( D ) The changes in mRNA expression levels of CTSS, TNF-α, RANTES, and IP-10 at 24 hpi were assessed by RT-qPCR. Data are presented as mean ± SD. Two to three independent experimental repeats were conducted for each assay. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, non-significant.

Article Snippet: For TNF-α treatment, human TNF-α protein (MedChem Express, USA HY-P7058) was added at 100 ng/mL 1.5 hpi.

Techniques: Virus, Expressing, Transfection, Infection, Western Blot, Gene Expression, Quantitative RT-PCR

Journal: Journal of Inflammation Research

Article Title: Exploring the Potential Value of Lactylation and Macrophage Polarization-Related Genes as Biomarkers for TNF-α Inhibitor Response in Inflammatory Bowel Disease

doi: 10.2147/JIR.S556906

Figure Lengend Snippet: Establishment of animal models. ( A ) Colon tissue. ( B ) Representative histological images of intestinal specimens. ( C ) Body weight and weight ratio (Day7/Day0). ( D ) DAI scores (Day7) and colon mucosa damage index. ( E ) RT-PCR of TNF-α expression levels. ( F ) RT-PCR of IL-6 expression levels. ( G ) RT-PCR of IL-10 expression levels. *P< 0.05, ***P<0.001, ****P<0.0001, ns, P> 0.05 (not significant).

Article Snippet: The TNF-α inhibitor group and the lactate combined TNF-α inhibitor group received 10 mg/kg of TNF-α inhibitor (MedChemExpress, 170277–31-3, USA) via intraperitoneal injection three times per week after the TNBS enema.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Journal of Inflammation Research

Article Title: Exploring the Potential Value of Lactylation and Macrophage Polarization-Related Genes as Biomarkers for TNF-α Inhibitor Response in Inflammatory Bowel Disease

doi: 10.2147/JIR.S556906

Figure Lengend Snippet: Effects of Lactate on TNF-α inhibitor Response. ( A ) RT-PCR of TNF-α expression levels. ( B ) RT-PCR of IL-6 expression levels. ( C ) RT-PCR of IL-10 expression levels. ( D ) Lactylation modifications. ( E ) Lactylation modifications relative level, Histone H3 served as loading control. *P< 0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: The TNF-α inhibitor group and the lactate combined TNF-α inhibitor group received 10 mg/kg of TNF-α inhibitor (MedChemExpress, 170277–31-3, USA) via intraperitoneal injection three times per week after the TNBS enema.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Control

Journal: Journal of Inflammation Research

Article Title: Exploring the Potential Value of Lactylation and Macrophage Polarization-Related Genes as Biomarkers for TNF-α Inhibitor Response in Inflammatory Bowel Disease

doi: 10.2147/JIR.S556906

Figure Lengend Snippet: Establishment of animal models. ( A ) Colon tissue. ( B ) Representative histological images of intestinal specimens. ( C ) Body weight and weight ratio (Day7/Day0). ( D ) DAI scores (Day7) and colon mucosa damage index. ( E ) RT-PCR of TNF-α expression levels. ( F ) RT-PCR of IL-6 expression levels. ( G ) RT-PCR of IL-10 expression levels. *P< 0.05, ***P<0.001, ****P<0.0001, ns, P> 0.05 (not significant).

Article Snippet: The TNF-α inhibitor group and the lactate combined TNF-α inhibitor group received 10 mg/kg of TNF-α inhibitor (MedChemExpress, 170277–31-3, USA) via intraperitoneal injection three times per week after the TNBS enema.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Journal of Inflammation Research

Article Title: Exploring the Potential Value of Lactylation and Macrophage Polarization-Related Genes as Biomarkers for TNF-α Inhibitor Response in Inflammatory Bowel Disease

doi: 10.2147/JIR.S556906

Figure Lengend Snippet: Effects of Lactate on TNF-α inhibitor Response. ( A ) RT-PCR of TNF-α expression levels. ( B ) RT-PCR of IL-6 expression levels. ( C ) RT-PCR of IL-10 expression levels. ( D ) Lactylation modifications. ( E ) Lactylation modifications relative level, Histone H3 served as loading control. *P< 0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: The TNF-α inhibitor group and the lactate combined TNF-α inhibitor group received 10 mg/kg of TNF-α inhibitor (MedChemExpress, 170277–31-3, USA) via intraperitoneal injection three times per week after the TNBS enema.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Control

Journal: Journal of Inflammation Research

Article Title: Exploring the Potential Value of Lactylation and Macrophage Polarization-Related Genes as Biomarkers for TNF-α Inhibitor Response in Inflammatory Bowel Disease

doi: 10.2147/JIR.S556906

Figure Lengend Snippet: Establishment of animal models. ( A ) Colon tissue. ( B ) Representative histological images of intestinal specimens. ( C ) Body weight and weight ratio (Day7/Day0). ( D ) DAI scores (Day7) and colon mucosa damage index. ( E ) RT-PCR of TNF-α expression levels. ( F ) RT-PCR of IL-6 expression levels. ( G ) RT-PCR of IL-10 expression levels. *P< 0.05, ***P<0.001, ****P<0.0001, ns, P> 0.05 (not significant).

Article Snippet: The TNF-α inhibitor group and the lactate combined TNF-α inhibitor group received 10 mg/kg of TNF-α inhibitor (MedChemExpress, 170277–31-3, USA) via intraperitoneal injection three times per week after the TNBS enema.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Journal of Inflammation Research

Article Title: Exploring the Potential Value of Lactylation and Macrophage Polarization-Related Genes as Biomarkers for TNF-α Inhibitor Response in Inflammatory Bowel Disease

doi: 10.2147/JIR.S556906

Figure Lengend Snippet: Effects of Lactate on TNF-α inhibitor Response. ( A ) RT-PCR of TNF-α expression levels. ( B ) RT-PCR of IL-6 expression levels. ( C ) RT-PCR of IL-10 expression levels. ( D ) Lactylation modifications. ( E ) Lactylation modifications relative level, Histone H3 served as loading control. *P< 0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: The TNF-α inhibitor group and the lactate combined TNF-α inhibitor group received 10 mg/kg of TNF-α inhibitor (MedChemExpress, 170277–31-3, USA) via intraperitoneal injection three times per week after the TNBS enema.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Control