Journal: BMC Gastroenterology

Article Title: Inhibition of TLR4 enhances oxaliplatin chemotherapy sensitivity in esophageal squamous cell carcinoma by suppressing inflammation and glycolysis

doi: 10.1186/s12876-026-04663-2

Figure Lengend Snippet: OXA activated TLR4 signaling in ESCC cells. The cells were treated by OXA (25 µM) for 24 h. A , B . The mRNA expressions of TLR4 and MYD88 in ESCC cell lines and normal esophageal cells were detected by qRT-PCR. C . ICC showed the immunocytochemical activity of NF-κB p65, p-NF-κB p65, COX-2, and MYD88. Scale bar: 50 μm. D . qRT-PCR analysis of mRNA levels of IL-1β, IL-6, COX-2, CXCL5, and CXCL8. *p < 0.05, **p < 0.01, ***p < 0.001 vs. Control group

Article Snippet: Serum concentrations of IL-6 (JL20268, Jonin) and IL-1β (KE10003, Proteintech) were quantified using commercial ELISA kits according to manufacturers’ protocols.

Techniques: Quantitative RT-PCR, Activity Assay, Control

Journal: BMC Gastroenterology

Article Title: Inhibition of TLR4 enhances oxaliplatin chemotherapy sensitivity in esophageal squamous cell carcinoma by suppressing inflammation and glycolysis

doi: 10.1186/s12876-026-04663-2

Figure Lengend Snippet: TLR4 knockout enhances the sensitivity of OXA chemotherapy in ESCC in vivo. A . Number of tumors. B . HE staining of esophageal epithelial tissues. Scale bar: 50 μm. C . Body weight of mice. D , E . the levels of serum IL-1β and IL-6 in mice from different groups were detected by ELISA. F . The immunohistochemical activities of PCNA, CK14, Cyclin D1, COX-2, S100A8 and S100A9 in the esophageal tissue were detected. Scale bar: 100 μm. G , H . The mRNA levels of inflammatory cytokines and glycolysis-related proteins in esophageal tissue were detected by qRT-PCR. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. WT group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. WT + 4NQO + OXA group

Article Snippet: Serum concentrations of IL-6 (JL20268, Jonin) and IL-1β (KE10003, Proteintech) were quantified using commercial ELISA kits according to manufacturers’ protocols.

Techniques: Knock-Out, In Vivo, Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Quantitative RT-PCR

Journal: BMC Gastroenterology

Article Title: Inhibition of TLR4 enhances oxaliplatin chemotherapy sensitivity in esophageal squamous cell carcinoma by suppressing inflammation and glycolysis

doi: 10.1186/s12876-026-04663-2

Figure Lengend Snippet: Schematic diagram illustrating the mechanism by which TLR4 inhibition enhances oxaliplatin (OXA) chemosensitivity in esophageal squamous cell carcinoma (ESCC). OXA treatment upregulates TLR4 and its downstream adaptor protein MYD88, which activates the phosphorylation of NF-κB p65. This activation drives two parallel pathways: (1) the inflammatory response, characterized by the upregulation of pro-inflammatory factors such as IL-6, COX-2, and CXCL5; (2) the glycolytic metabolic reprogramming, mediated by the HIF-1α/GLUT1 axis and enhanced expression of glycolytic enzymes including PFKM and LDHB. These two pathways synergistically promote ESCC cell proliferation, migration, and invasion, ultimately reducing OXA chemosensitivity. Inhibition of TLR4 (via genetic knockout, shRNA knockdown, or pharmacological inhibitor TAK-242) or its downstream mediator MYD88 (via shRNA knockdown or inhibitor ST2825) blocks NF-κB p65 phosphorylation, thereby suppressing both the inflammatory response and glycolytic activity. This dual inhibition disrupts the adaptive survival mechanisms of ESCC cells, potentiating the anti-tumor efficacy of OXA

Article Snippet: Serum concentrations of IL-6 (JL20268, Jonin) and IL-1β (KE10003, Proteintech) were quantified using commercial ELISA kits according to manufacturers’ protocols.

Techniques: Inhibition, Phospho-proteomics, Activation Assay, Expressing, Migration, Knock-Out, shRNA, Knockdown, Activity Assay

Journal: Frontiers in Pharmacology

Article Title: Radix Pseudostellaria polysaccharides alleviate sepsis-induced liver injury by modulating the gut microbiota via the TLR4/NF-κB pathway

doi: 10.3389/fphar.2025.1658147

Figure Lengend Snippet: Radix Pseudostellariae polysaccharides (RPPS) attenuate CLP-induced liver inflammation and injury in mice. (A) The experimental design and procedures for RPPS administration. (B) Survival rate. (C) MSS in the NS + CLP and RP + CLP groups. (D) Liver morphology. (E) Histological representation of liver sections using hematoxylin and eosin (H,E) staining (×100 and ×200 magnification, n = 10). (F) Immunohistochemical staining of CD68 (×100 and ×200 magnification, n = 6). (G) Representative transmission electron microscopy images of hepatocytes. Bar = 1 μm (n = 6–8). (H–J) Alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin (TBIL) levels. (K) Histopathological score from H&E sections. (L) The rate of CD68-positive cell density in the liver. (M–O) Levels of TNF-α, IL-1β, and IL-6 in liver tissue. Data are presented as mean ± standard error of mean. A one-way analysis of variance with Tukey’s analysis was used for multiple comparisons. Significance levels are represented as: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, with “ns” indicating “not significant.”

Article Snippet: Subsequently, slides were blocked with 10% goat serum (SL038, Solarbio, China) at room temperature for 45 min. Primary antibodies against CD68: 1:200, 28058-1-AP; TNF-α: 1:200, 26405-1-AP; IL-1β: 1:200, 26048-1-AP, all sourced from Proteintech, were incubated at 4 °C for 16 h. Endogenous peroxidase activity was blocked by 20-min incubation in 3% hydrogen peroxide.

Techniques: Staining, Immunohistochemical staining, Transmission Assay, Electron Microscopy

Journal: Frontiers in Pharmacology

Article Title: Radix Pseudostellaria polysaccharides alleviate sepsis-induced liver injury by modulating the gut microbiota via the TLR4/NF-κB pathway

doi: 10.3389/fphar.2025.1658147

Figure Lengend Snippet: The beneficial effects of Radix Pseudostellariae polysaccharides (RPPS) on sepsis progression are associated with the TLR4/NF-κB pathway. (A) Immunofluorescent staining of Toll-like receptor 4 (TLR4) in liver tissues. (×200 magnification, n = 3). (B) Quantification of the percentage of TLR4-positive cells. (C) Immunofluorescent staining of p-p65 in liver tissues. (D) Quantification of the percentage of p-p65-positive cells. (Bar = 50 μm, n = 3). (E) Representative Western blot images of the NF-κB pathway. (F–J) Expression and quantification of MyD88, IKK α/β, NF-κB (p65), and pNF-κB (pp65) proteins using Western blot (n = 3). (K) Immunohistochemical staining for IL-1β and TNFα (×100 and ×200 magnification). (L,M) Percentage of IL-1β and TNFα positive area in liver tissue (n = 6). Data are expressed as the mean ± standard error of mean, and the significance of the differences between groups was assessed using one-way analysis of variance (Tukey’s test). * p < 0.05, ** p < 0.05.

Article Snippet: Subsequently, slides were blocked with 10% goat serum (SL038, Solarbio, China) at room temperature for 45 min. Primary antibodies against CD68: 1:200, 28058-1-AP; TNF-α: 1:200, 26405-1-AP; IL-1β: 1:200, 26048-1-AP, all sourced from Proteintech, were incubated at 4 °C for 16 h. Endogenous peroxidase activity was blocked by 20-min incubation in 3% hydrogen peroxide.

Techniques: Staining, Western Blot, Expressing, Immunohistochemical staining

Journal: JCI Insight

Article Title: FGF receptors mediate cellular senescence in the cystic fibrosis airway epithelium

doi: 10.1172/jci.insight.174888

Figure Lengend Snippet: Single-cell RNA transcriptome of control and CF epithelial cells (National Center for Biotechnology Gene Expression Omnibus GSE150674) from 19 control and 19 CF donor lungs from donors with end-stage CF lung disease and healthy controls, CF ΔF508 homozygous patients ( n = 10,131 cells and n = 8 donors) and healthy donors ( n = 23,119 cells and n = 19) were separated from the whole data set and analyzed using BBrowser3 to generate UMAPs and violin plots showing senescence scores from 3 separate gene databases: ( A and B ) CellAge database of senescence inducing genes (416 genes), ( C and D ) SenMayo (124 genes), and ( E and F ) cellular senescence markers used in this study ( CDKN1A , CDKN2A , BCL2 , BCL2L1 , IL6 , IL1B , IL6 , and GLB1 ). ( G ) Senescence scores using the same 3 gene databases listed above but separated into violin plots looking at senescence scores based on major epithelial cell types: basal, ciliated, and secretory. Statistical analysis was done using unpaired Student’s t test or 1-way ANOVA shown with **** P < 0.0001.

Article Snippet: Real-time quantitative PCR was performed with the following TaqMan probes: IL-6 Hs00174131, IL-1β Hs01555410, CXCL8 Hs00174103, BCL2L1 Hs00236329_m1 GAPDH (4333764F), FGFR1 Rn01478647, FGFR2 Rn01269940, FGFR3 Rn00584799, FGFR4 Rn01441815, BCL2 Rn99999125, p21 Rn00589996, p16 Rn00580664, IL-8 Rn00586403, IL-1β Rn00580432, IL-6 Rn01410330, BCL2L1 Rn06267811_g1, GAPDH Rn01775763 (Invitrogen, Thermo Fisher Scientific).

Techniques: Control, Gene Expression

Journal: JCI Insight

Article Title: FGF receptors mediate cellular senescence in the cystic fibrosis airway epithelium

doi: 10.1172/jci.insight.174888

Figure Lengend Snippet: ( A ) Representative immunoblot images and bar graphs showing densitometric analyses for p16, p21, and BCL-xL in CF ΔF508 and non-CF donors at ALI ( N = 5). ( B ) Representative images for SA-β-gal staining using brightfield imaging of the same CF ΔF508 and non-CF donor ALI cultures including quantification of β-gal staining using the ratio of SA-β-gal–positive cells per brightfield by ImageJ (NIH) ( N = 3); arrows show β-gal–positive cells (scale bar = 100 μm, original magnification, ×40). ( C ) Bar graphs demonstrating relative mRNA levels of SASP ( IL1B , IL6 , and IL8 ) markers normalized to GAPDH. ( D ) Bar graphs indicating relative mRNA levels of FGFR1–4 normalized to GAPDH in the same 2 groups. Statistical analysis was done using unpaired Student’s t test showing means ± SEM with * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 from 3–5 different donors per group with experiments repeated 3 times.

Article Snippet: Real-time quantitative PCR was performed with the following TaqMan probes: IL-6 Hs00174131, IL-1β Hs01555410, CXCL8 Hs00174103, BCL2L1 Hs00236329_m1 GAPDH (4333764F), FGFR1 Rn01478647, FGFR2 Rn01269940, FGFR3 Rn00584799, FGFR4 Rn01441815, BCL2 Rn99999125, p21 Rn00589996, p16 Rn00580664, IL-8 Rn00586403, IL-1β Rn00580432, IL-6 Rn01410330, BCL2L1 Rn06267811_g1, GAPDH Rn01775763 (Invitrogen, Thermo Fisher Scientific).

Techniques: Western Blot, Staining, Imaging

Journal: JCI Insight

Article Title: FGF receptors mediate cellular senescence in the cystic fibrosis airway epithelium

doi: 10.1172/jci.insight.174888

Figure Lengend Snippet: ( A ) Immunohistochemical staining for p16, p21, and BCL-xL in Cftr –/– rat lung tissue compared to controls demonstrating an increased signal in the bronchial epithelium (scale bar = 100 μm, original magnification, ×10; for insets, ×20). ( B ) SA-β-gal stain and nuclear counterstain (DAPI) in lung tissue from Cftr –/– rats and littermate controls; arrows indicate areas of airway epithelial β-gal staining (scale bar = 100 μm, original magnification, ×40). ( C ) Relative mRNA levels of Cdkn2a (p16), Cdkn1a (p21), Bcl2l1 , and SASP markers ( Il1b , Il6 , and Cxcl2 [IL-8]) normalized to GAPDH, from total lung tissue of control and Cftr –/– rats. Statistical analysis was done using unpaired Student’s t test showing means ± SEM with * P < 0.05, ** P < 0.01 with n = 5–10 rats per group and experiments done in triplicates.

Article Snippet: Real-time quantitative PCR was performed with the following TaqMan probes: IL-6 Hs00174131, IL-1β Hs01555410, CXCL8 Hs00174103, BCL2L1 Hs00236329_m1 GAPDH (4333764F), FGFR1 Rn01478647, FGFR2 Rn01269940, FGFR3 Rn00584799, FGFR4 Rn01441815, BCL2 Rn99999125, p21 Rn00589996, p16 Rn00580664, IL-8 Rn00586403, IL-1β Rn00580432, IL-6 Rn01410330, BCL2L1 Rn06267811_g1, GAPDH Rn01775763 (Invitrogen, Thermo Fisher Scientific).

Techniques: Immunohistochemical staining, Staining, Control

Journal: JCI Insight

Article Title: FGF receptors mediate cellular senescence in the cystic fibrosis airway epithelium

doi: 10.1172/jci.insight.174888

Figure Lengend Snippet: ( A ) Immunohistochemistry of p16 and p21 from ex vivo Cftr –/– rat tracheae treated with BLU9931 at 0.1 M for 24 hours compared with vehicle-treated “control” Cftr –/– rat tracheae. ( B ) mRNA levels of Cdkn2a (p16), Cdkn1a (p21), Bcl2 , and Bcl2l1 (BCL-xL) and SASP markers ( C ) Il1b , Il6 , and Cxcl2 (IL8) from the ex vivo Cftr –/– rat tracheae +/– BLU9931. ( D ) Representative µOCT images (ep, epithelial layer; lp, lamina propria) and ( E ) bar graphs showing µOCT quantification of periciliary liquid depth (PCL), mucociliary transport (MCT), ciliary beat frequency (CBF), and airway surface liquid (ASL) depth both from BLU9931 and vehicle control treated ex vivo Cftr –/– rat tracheae. Statistical analysis was done using unpaired Student’s t test showing means ± SEM with * P < 0.05, ** P < 0.01, with n = 3–4 rat trachea per group.

Article Snippet: Real-time quantitative PCR was performed with the following TaqMan probes: IL-6 Hs00174131, IL-1β Hs01555410, CXCL8 Hs00174103, BCL2L1 Hs00236329_m1 GAPDH (4333764F), FGFR1 Rn01478647, FGFR2 Rn01269940, FGFR3 Rn00584799, FGFR4 Rn01441815, BCL2 Rn99999125, p21 Rn00589996, p16 Rn00580664, IL-8 Rn00586403, IL-1β Rn00580432, IL-6 Rn01410330, BCL2L1 Rn06267811_g1, GAPDH Rn01775763 (Invitrogen, Thermo Fisher Scientific).

Techniques: Immunohistochemistry, Ex Vivo, Control

Journal: JCI Insight

Article Title: FGF receptors mediate cellular senescence in the cystic fibrosis airway epithelium

doi: 10.1172/jci.insight.174888

Figure Lengend Snippet: ( A ) Immunohistochemistry of Cdkn2a (p16) and Cdkn1a (p21) of Cftr –/– rat tracheae, which were treated with 100 nM dasatinib and 2 μM quercetin (D+Q) for 24 hours, compared with vehicle-treated Cftr –/– rat tracheae. Scale bar is 100 µm. ( B ) Representative µOCT images of the different Cftr –/– rat trachea groups (ep, epithelial layer; lp, lamina propria), including representative images to assess mucociliary transport (MCT). ( C ) Bar graphs showing quantification of all regions of interest from µOCT images for assessment of ASL, MCT, CBF, and PCL from the vehicle- and D+Q-treated groups. ( D ) mRNA levels of SASP markers ( Il1b , Il6 , and Cxcl2 [IL-8]) along with ( E ) senescence markers Cdkn2a (p16), Cdkn1a (p21), Bcl2 , and Bcl2l1 (BCL-xL) from Cftr –/– rat tracheae ± D+Q. Statistical analysis was done using unpaired Student’s t test showing means ± SEM with * P < 0.05, ** P < 0.01 with n = 3–4 rat tracheae per group.

Article Snippet: Real-time quantitative PCR was performed with the following TaqMan probes: IL-6 Hs00174131, IL-1β Hs01555410, CXCL8 Hs00174103, BCL2L1 Hs00236329_m1 GAPDH (4333764F), FGFR1 Rn01478647, FGFR2 Rn01269940, FGFR3 Rn00584799, FGFR4 Rn01441815, BCL2 Rn99999125, p21 Rn00589996, p16 Rn00580664, IL-8 Rn00586403, IL-1β Rn00580432, IL-6 Rn01410330, BCL2L1 Rn06267811_g1, GAPDH Rn01775763 (Invitrogen, Thermo Fisher Scientific).

Techniques: Immunohistochemistry

Journal: JCI Insight

Article Title: FGF receptors mediate cellular senescence in the cystic fibrosis airway epithelium

doi: 10.1172/jci.insight.174888

Figure Lengend Snippet: ( A ) Representative µOCT images and bar graph of CF primary human bronchial epithelial cells ( n = 3 donors per group) treated with VX-661/VX-445/VX-770 (ETI) for 72 hours showing a significant increase in ASL depth in CF cells treated with ETI; yellow line is ASL depth. ep, epithelial layer; F, filter. ( B ) Representative immunoblots and densitometric analysis showing no significant difference in protein expression of cellular senescence markers p16, p21, and BCL-xL in CF primary human bronchial epithelial cells treated with ETI for 72 hours when compared with untreated CF primary human bronchial epithelial cells ( n = 4 donors per group). ( C ) Relative expression of SASP markers ( Il1b , Il6 , and Cxcl2 [IL-8]) from CF primary human bronchial epithelial cells demonstrates no significant change when compared with untreated CF primary human bronchial epithelial cells. ( D ) Immunoblots and densitometric analysis of p21 and BCL-xL in hG551D rats and hG551D rats treated with VX-770 for 14 days (hG551d = 3 rats, hG551d+VX-770 = 5 rats). ( E ) Relative expression of SASP markers Il1b , Il6 , and Cxcl2 (IL-8) in hG551D rats and hG551D rats treated with VX-770 for 14 days (hG551d = 3 rats, hG551d+VX-770 = 5 rats). Statistical analysis was done using unpaired Student’s t test showing means ± SEM with * P < 0.05.

Article Snippet: Real-time quantitative PCR was performed with the following TaqMan probes: IL-6 Hs00174131, IL-1β Hs01555410, CXCL8 Hs00174103, BCL2L1 Hs00236329_m1 GAPDH (4333764F), FGFR1 Rn01478647, FGFR2 Rn01269940, FGFR3 Rn00584799, FGFR4 Rn01441815, BCL2 Rn99999125, p21 Rn00589996, p16 Rn00580664, IL-8 Rn00586403, IL-1β Rn00580432, IL-6 Rn01410330, BCL2L1 Rn06267811_g1, GAPDH Rn01775763 (Invitrogen, Thermo Fisher Scientific).

Techniques: Western Blot, Expressing

Journal: Japanese Clinical Medicine

Article Title: Dynamic Change in Cells Expressing IL-1β in Rat Hippocampus after Status Epilepticus

doi: 10.4137/jcm.s13738

Figure Lengend Snippet: Figure 2. IL-1b immunoreactivity in the hippocampus of control rat without SE and after SE. (A, D, G, J, M) Expression of IL-1b at control rat hippocampus. (A) Expression of IL-1b at the hippocampus of control rat without SE in low-power magnification. Scale bar = 1 mm. (D) CA1 region of control rat in intermediate power magnification. (G) CA3 region of control rat in intermediate power magnification. D, G; scale bar = 100 μm. (J) CA1 region of control rat in high-power magnification. (M) CA3 region of control rat in high-power magnification. J, M; scale bar = 50 μm. Expression of IL-1b was faintly observed in the pyramidal cells at CA1 (D, J) and CA3 (G, M) regions. (B, E, H, K, N) Expression of IL-1b at day 1 after SE. (B) Expression of IL-1b at the hippocampus at day 1 after SE in low power magnification. Scale bar = 1 mm. (E) CA1 region at day 1 after SE in intermediate power magnification. (H) CA3 region at day 1 after SE in intermediate power magnification. E, H; scale bar = 100 μm. (K) CA1 region at day 1 in high power magnification. (N) CA3 region at day 1 after SE in high power magnification. IL-1b expression increased transiently in the cytoplasm of the remaining pyramidal cells in the CA3 region beginning on day 1 after SE (H, N). Expression of IL-1b in the cytoplasm of pyramidal cells at CA3 was greater than that at CA1 on day 1 after SE (E, H, K, N; Table 1). K, N; scale bar = 50 μm. (C, F, I, L, O) Expression of IL-1b at day 21 after SE. (B) Expression of IL-1b at the hippocampus at day 21 after SE in low-power magnification. Scale bar = 1 mm. (E) CA1 region at day 21 after SE in intermediate power magnification. (H) CA3 region at day 21 after SE in intermediate power magnification. F, I; scale bar = 100 μm. (F) CA1 region at day 21 in high power magnification. (N) CA3 region at day 21 after SE in high power magnification. Reactive astrocyte-like cells emerging in CA1 showed IL-1b immunoreactivity on day 7. This immunointensity increased in proportion to progressive hypertrophy until day 21. L, O; scale bar = 50 μm.

Article Snippet: For the immunohistochemical study, a commercially available rabbit polyclonal anti-IL-1b antibody (diluted 1:500; sc-1251, Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used.

Techniques: Control, Expressing

Journal: Japanese Clinical Medicine

Article Title: Dynamic Change in Cells Expressing IL-1β in Rat Hippocampus after Status Epilepticus

doi: 10.4137/jcm.s13738

Figure Lengend Snippet: Figure 3. Double-label immunofluorescence staining of IL-1b and GFAP. Co-localization of IL-1b (red, A) and GFAP (green, B) is shown with an immunofluorescence method. Co-localization is visualized in yellow in the merged image (C). Double-label fluorescent immunohistochemistry clarified that reactive astrocytes expressed IL-1b. Scale bar = 50 μm.

Article Snippet: For the immunohistochemical study, a commercially available rabbit polyclonal anti-IL-1b antibody (diluted 1:500; sc-1251, Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used.

Techniques: Immunofluorescence, Staining, Immunohistochemistry

Journal: Japanese Clinical Medicine

Article Title: Dynamic Change in Cells Expressing IL-1β in Rat Hippocampus after Status Epilepticus

doi: 10.4137/jcm.s13738

Figure Lengend Snippet: Figure 4. Total IL-1b expression in the hippocampus after SE. The total expression level IL-1b in the hippocampus measured by using Luminex technology was significantly elevated from day 1 after SE and maintained till day 21 (P 0.01). Asterisks (*) and daggers (†) indicate significant differences (P 0.05 and P 0.005, respectively) from the value for the control group.

Article Snippet: For the immunohistochemical study, a commercially available rabbit polyclonal anti-IL-1b antibody (diluted 1:500; sc-1251, Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used.

Techniques: Expressing, Luminex, Control

Journal: Journal of Translational Medicine

Article Title: 18-β-glycyrrhetinic acid facilitates nuclear-mitochondrial communications to alleviate oxidative stress through HMGB1-cGAS-Mul1 axis in tendinopathy

doi: 10.1186/s12967-026-08091-4

Figure Lengend Snippet: Oxidative stress levels are increased in patients with tendinopathy. The tendons of normal clinical samples and tendinopathy patients were collected and tested. ( A ) Proteomics was used for detection, and the results were subjected to GO analysis for Biological Process, Cellular Component and Molecular Function of proteins. ( B ) The protein expression of PTGS2 in tendon tissues was assessed by immunohistochemical staining (Scale bars: 20×, 50 μm). ( C-D ) Uniform manifold approximation and projection (UMAP) was used to visualize nuclear clusters, which are coloured and labelled according to cell identities, in normal (left panel) and injured (right panel) tendon tissues. ( E-F ) Proportion of cell types in normal and injured tendon tissues. Each cell type is colour-coded. ( G ) Immunofluorescence staining identifying TSCs (Red: SCX + ; Green: CD146 + ). ( H ) The superoxide levels in TSCs (SCX + ) and macrophages (CD68 + ) were detected by DHE staining

Article Snippet: The anti-TNMD antibody (#A17753), anti-IL-6 antibody (#A22222), anti-IL-1β antibody (#A16288), anti-TBK1 antibody (#A3458), anti-P-IRF3-S386 antibody (#AP0857) and anti-P-TBK1-S172 antibody (#AP1026) were purchased from ABclonal Technology.

Techniques: Expressing, Immunohistochemical staining, Staining, Immunofluorescence

Journal: Journal of Translational Medicine

Article Title: 18-β-glycyrrhetinic acid facilitates nuclear-mitochondrial communications to alleviate oxidative stress through HMGB1-cGAS-Mul1 axis in tendinopathy

doi: 10.1186/s12967-026-08091-4

Figure Lengend Snippet: GA attenuates oxidative stress by inhibiting inflammation to alleviate tendinopathy. In vivo, Wistar rats were divided into 6 groups: vehicle, ciprofloxacin, ciprofloxacin + 25 mg/kg GA, ciprofloxacin + 50 mg/kg GA, ciprofloxacin + 100 mg/kg GA and positive control (five rats in each group). ( A ) H&E and Masson staining were used to observe collagen organization (blue: collagen fibres; red: muscle fibres); Sirius red staining was used to distinguish between type I and type III collagen (scale bars: 10×, 100 μm); ( B ) The expression of collagen I/III was determined by immunofluorescence staining (scale bars: 10×, 100 μm); ( C ) The protein expression of PTGS2 in rat tendon tissues was assessed by immunohistochemical staining (scale bars: 10×, 100 μm). ( D-E ) Biological process and protein function enrichment analysis of the TMT proteomics data. ( F-H ) The serum levels of MDA, GSH and IL-1β were determined with the relevant kits; ( n = 5, data are presented as mean ± SD, ** p < 0.01, *** p < 0.001 vs. vehicle group; ## p < 0.01, ### p < 0.001 vs. Ciprofloxacin group)

Article Snippet: The anti-TNMD antibody (#A17753), anti-IL-6 antibody (#A22222), anti-IL-1β antibody (#A16288), anti-TBK1 antibody (#A3458), anti-P-IRF3-S386 antibody (#AP0857) and anti-P-TBK1-S172 antibody (#AP1026) were purchased from ABclonal Technology.

Techniques: In Vivo, Positive Control, Staining, Expressing, Immunofluorescence, Immunohistochemical staining

Journal: Journal of Translational Medicine

Article Title: 18-β-glycyrrhetinic acid facilitates nuclear-mitochondrial communications to alleviate oxidative stress through HMGB1-cGAS-Mul1 axis in tendinopathy

doi: 10.1186/s12967-026-08091-4

Figure Lengend Snippet: GA increases TSC viability by inhibiting oxidative stress and inflammatory responses. In vitro, TSCs were pretreated with different concentrations of GA for 12 h and then subjected to the prescribed concentration of H 2 O 2 (100 µM) for 24 h. ( A-B ) Cell viability was assessed by MTT assay; ( C ) Cell viability was assessed by CCK-8 assay; ( D-E ) The levels of GSH and MDA were determined with the relevant kits; ( F ) The content of ROS in TSCs was measured by flow cytometry using DCFH-DA probe; ( G-H ) Western blotting and qPCR were used to assess the protein and gene expression of inflammatory cytokines, including IL-1β, IL-6, and IL-8; ( I ) Tenomodulin, Collagen I and Collagen III protein levels were assessed via western blotting. ( n = 3, data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H₂O₂ group)

Article Snippet: The anti-TNMD antibody (#A17753), anti-IL-6 antibody (#A22222), anti-IL-1β antibody (#A16288), anti-TBK1 antibody (#A3458), anti-P-IRF3-S386 antibody (#AP0857) and anti-P-TBK1-S172 antibody (#AP1026) were purchased from ABclonal Technology.

Techniques: In Vitro, Concentration Assay, MTT Assay, CCK-8 Assay, Flow Cytometry, Western Blot, Gene Expression, Control

Journal: Journal of Translational Medicine

Article Title: 18-β-glycyrrhetinic acid facilitates nuclear-mitochondrial communications to alleviate oxidative stress through HMGB1-cGAS-Mul1 axis in tendinopathy

doi: 10.1186/s12967-026-08091-4

Figure Lengend Snippet: GA inhibits the cGAS-STING-NLRP3 axis to alleviate H 2 O 2 -induced TSC damage. TSCs were pretreated with the cGAS plasmid or GA at different concentrations for 12 h, and then exposed to 100 µM H 2 O 2 (freshly diluted in media) for 24 h. ( A-B ) Protein levels of the cGAS-STING axis were evaluated by western blotting; ( C ) The mRNA levels of IFN-β were measured by qPCR; ( D ) An IFN-β ELISA kit was used to detect the content of IFN-β in TSCs; ( E ) The NLRP3 and ASC protein levels were evaluated by western blotting; ( F ) Western blotting was used to verify the overexpression efficiency of the cGAS plasmid; ( G ) The IFN-β ELISA kit was used to detect the content of IFN-β in TSCs; ( H ) Western blotting was used to detect the protein expression of inflammatory cytokines, including IL-1β and IL-6. ( I ) NLRP3 and ASC protein levels were evaluated by western blotting; ( J-K ) The corresponding kits were used to measure the levels of oxidative stress in TSCs; ( n = 3, data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group, # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H₂O₂ group, & p < 0.05, && p < 0.01 vs. H₂O₂ + GA group)

Article Snippet: The anti-TNMD antibody (#A17753), anti-IL-6 antibody (#A22222), anti-IL-1β antibody (#A16288), anti-TBK1 antibody (#A3458), anti-P-IRF3-S386 antibody (#AP0857) and anti-P-TBK1-S172 antibody (#AP1026) were purchased from ABclonal Technology.

Techniques: Plasmid Preparation, Western Blot, Enzyme-linked Immunosorbent Assay, Over Expression, Expressing, Control

Journal: Journal of Translational Medicine

Article Title: 18-β-glycyrrhetinic acid facilitates nuclear-mitochondrial communications to alleviate oxidative stress through HMGB1-cGAS-Mul1 axis in tendinopathy

doi: 10.1186/s12967-026-08091-4

Figure Lengend Snippet: GA inhibits the expression of cGAS by promoting its ubiquitination-mediated degradation. ( A ) The cGAMP ELISA kit was used to assess the enzyme activity of cGAS; ( B ) qPCR was used to detect the mRNA expression of cGAS under different GA concentrations; ( C ) TSCs were treated with cycloheximide (CHX, 20 µg/ml) in the presence or absence of GA (5 µM), and cGAS levels were analyzed by western blot at the indicated time points over 8 h; ( D-E ) TSCs were pretreated with 3-MA (5 µM), CHX (20 µg/ml) or MG-132 (1.25 µM) for 4 h and then exposed to 100 µM H 2 O 2 (freshly diluted in media) for 24 h, the protein expression of cGAS was detected by western blotting; ( F ) Ubiquitination of cGAS was analyzed by immunoprecipitation (IP) followed by immunoblotting (IB); ( G ) Ubibrowser ( http://ubibrowser.ncpsb.org/ ) was used to predict the E3 ubiquitin ligase of cGAS in rats; ( H ) The binding of cGAS and predicted E3 ubiquitin ligase was detected by co-immunoprecipitation; ( I ) The colocalization of cGAS and TOMM20 was detected by immunofluorescence staining. ( n = 3, data are presented as mean ± SD, ** p < 0.01, *** p < 0.001 vs. control group; ## p < 0.01, ### p < 0.001 vs. H₂O₂ group; && p < 0.01, &&& p < 0.001 vs. H₂O₂+GA group)

Article Snippet: The anti-TNMD antibody (#A17753), anti-IL-6 antibody (#A22222), anti-IL-1β antibody (#A16288), anti-TBK1 antibody (#A3458), anti-P-IRF3-S386 antibody (#AP0857) and anti-P-TBK1-S172 antibody (#AP1026) were purchased from ABclonal Technology.

Techniques: Expressing, Ubiquitin Proteomics, Enzyme-linked Immunosorbent Assay, Activity Assay, Western Blot, Immunoprecipitation, Binding Assay, Immunofluorescence, Staining, Control

Journal: Journal of Translational Medicine

Article Title: 18-β-glycyrrhetinic acid facilitates nuclear-mitochondrial communications to alleviate oxidative stress through HMGB1-cGAS-Mul1 axis in tendinopathy

doi: 10.1186/s12967-026-08091-4

Figure Lengend Snippet: GA protects TSCs against oxidative stress via the HMGB1-cGAS-NLRP3 axis in tendinopathy. Wistar rats were divided into five groups (five rats per group): vehicle, ciprofloxacin, ciprofloxacin + 50 mg/kg GA, ciprofloxacin + 50 mg/kg GA + VA-Lip-Ctrl-Plasmid, and ciprofloxacin + 50 mg/kg GA + VA-Lip-cGAS-Plasmid. ( A-B ) Immunohistochemical detection for cGAS, HMGB1, NLRP3 and ASC in tendon tissues (scale bars: 10×, 100 μm); ( C ) H&E and Masson staining were used to observe the collagen organization of tendon tissues (blue indicates collagen fibres and red indicates muscle fibres); Sirius red staining was used to distinguish between type I and type III collagen (Scale bars: 10×, 100 μm); ( D ) The expression of collagen I/III were assessed by immunofluorescence staining (scale bars: 10×, 100 μm); ( E ) Immunohistochemical detection of the expression of NLRP3 and ASC (scale bars: 10×, 100 μm); ( F-H ) Serum levels of MDA and IL-1β were measured by commercial ELISA kits; ( I ) Immunofluorescence staining was used to detect the expression and colocalization of cGAS and HMGB1 in the tendon tissues of the rats (scale bars: 20×, 50 μm). ( J ) Immunofluorescence staining was used to detect the colocalization of cGAS and HMGB1 in the tendon tissue of clinical sample 1. ( n = 5, data are presented as mean ± SD, ** p < 0.01, *** p < 0.001 vs. vehicle group; ## p < 0.01, ### p < 0.001 vs. Ciprofloxacin group; && p < 0.01, &&& p < 0.001 vs. Ciprofloxacin + GA group)

Article Snippet: The anti-TNMD antibody (#A17753), anti-IL-6 antibody (#A22222), anti-IL-1β antibody (#A16288), anti-TBK1 antibody (#A3458), anti-P-IRF3-S386 antibody (#AP0857) and anti-P-TBK1-S172 antibody (#AP1026) were purchased from ABclonal Technology.

Techniques: Plasmid Preparation, Immunohistochemical staining, Staining, Expressing, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Molecular Oncology

Article Title: Plasminogen deficiency suppresses pancreatic ductal adenocarcinoma disease progression

doi: 10.1002/1878-0261.13552

Figure Lengend Snippet: Elimination of plasminogen reduces the accumulation of pro‐tumor immune cell populations in the KPC2 PDAC tumor microenvironment. Representative images ( left ) and quantification (as appropriate) of immunohistochemical staining ( right ) for (A) H&E, (B) CD3, (C) AIF‐1, (D) Ly6G, and (E) CD31 in tumor tissue harvested 3 weeks after orthotopic injection into Plg + ( n = 9) and Plg − ( n = 6) mice. Scale bar = 100 μm. Quantitative RT‐PCR analysis of tumor tissue harvested from Plg + ( n = 14) and Plg − ( n = 13) mice orthotopically injected with KPC2 cells for (F) Tnfa , (G) Il6 , (H) Il1b , (I) Ccl2 , and (J) Il10 . Data are presented as mean ± standard error of the mean and were analyzed by Mann–Whitney test with * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Quantitative RT‐qPCR was performed with the TaqManTM Gene Expression Master Mix (ThermoFisher Scientific cat# 4369016) using probes for Plg (Mm00447087_m1), Fga (Mm00802584_m1), Egf (Mm00438696_m1), Hgf (Mm01135184_m1), Fgf2 (Mm01285715_m1), Tgfb1 (Mm01178820_m1), Mmp1 (Mm00473485_m1), Mmp2 (Mm00439498_m1), Mmp9 (Mm00442991_m1), Mmp13 (Mm00439491_m1), Il1b (Mm00434228_m1), Il6 (Mm00446190_m1), Il10 (Mm01288386_m1), Cdh1 (Mm01247357_m1), Plgrkt (Mm00509491_g1), or S100a10 (Mm00501458_g1).

Techniques: Immunohistochemical staining, Staining, Injection, Quantitative RT-PCR, MANN-WHITNEY

Journal: The Journal of antimicrobial chemotherapy

Article Title: Induction of interleukin-1beta, tumour necrosis factor-alpha and apoptosis in mouse organs by amphotericin B is neutralized by conjugation with arabinogalactan.

doi: 10.1093/jac/dki090

Figure Lengend Snippet: Figure 3. Immunohistochemical staining of mouse kidneys for IL-1b demonstrating positively-stained tubular cells in mice treated with 4 mg/kg AMB-AG conjugate (a), compared with untreated mice in which no staining was demonstrated (b). The specific staining for IL-1b, originally demon- strated by a reddish-brown hue, is presented here in dark-black. Kidneys were excised 4 h post-injection. Magnification, 400.

Article Snippet: Goat polyclonal antibody against mouse IL-1b (R&D Systems, Inc., Minneapolis, MN, USA) that served as the primary detecting antibody was diluted in CAS-Blocker to 5mg/mL.

Techniques: Immunohistochemical staining, Staining, Injection

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: Significant Ups genes in the acetabular cartilage of the rats with DDH, as determined by mRNA-seq analysis. (A) Neonatal rats were subjected to straight-leg swaddling for 10 days. A total of 4 weeks later, (a) morphology, (b) H&E staining and (c) SO/FG staining were assessed in the hip joints of the rats. (B) Immunohistochemical staining and (E) semi-quantitative analysis of COL2A1 in the acetabular cartilage of the 4-week-old control rats and rats with DDH. (C) Relative gene expression levels of COL2A1, IL-1β and IL-6 in the acetabular cartilage of the 4-week-old control rats and rats with DDH, as determined by reverse transcription-quantitative PCR. (D) Levels of IL-6 in the supernatants of acetabular cartilage from the 4-week-old rats with DDH, as determined by ELISA. (F) Significant Ups in the acetabular cartilage of rats with DDH were identified by mRNA-seq analysis with the screening criteria of log 2 FC >4 and P.adj<0.01. Principal component analysis results and a heatmap of these significant Ups are shown. (G) GO enrichment analysis of significant Ups in the acetabular cartilage of the rats with DDH. Data are presented as the mean ± SD. *** P<0.001. (C-E) Representative results of four independent experiments are shown. Ups, upregulated genes; BP, biological process; CC, cellular component; COL2A1, type II collagen; DDH, developmental dysplasia of the hip; FC, fold change; GO, Gene Ontology; H&E, hematoxylin and eosin; IHC, immunohistochemistry; IL, interleukin; IOD, integrated optical density; MF, molecular function; mRNA-seq, mRNA sequencing; SO/FG, Safranin O/Fast Green.

Article Snippet: To assess the functional integrity of chondrocytes, the P0-P1 primary chondrocytes were treated with the pro-inflammatory cytokine IL-1β (10 ng/ml; cat. no. HY-P7097; MedChemExpress) with or without the HDAC inhibitor trichostatin A (TSA; 300 nM; cat. no. T129665; Shanghai Aladdin Biochemical Technology Co., Ltd.) for 24 h at room temperature ( , ).

Techniques: Staining, Immunohistochemical staining, Control, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Sequencing

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: DUSP26 expression is upregulated in IL-1β-induced chondrocytes. (A) Schematic diagram showing the experimental protocol in vitro . Primary chondrocytes isolated from 5-day-old rats were subjected to treatment with 10 ng/ml IL-1β. The protein and mRNA expression levels of DUSP26 in chondrocytes were detected by RT-qPCR and western blotting. (B) Rat DUSP26 overexpression adenovirus was constructed and used to infect chondrocytes. After 24 h, the protein and mRNA expression levels of DUSP26 in the chondrocytes were detected by RT-qPCR and western blotting. (C) Two adenovirus-mediated shRNAs against rat DUSP26 were constructed and used to infect chondrocytes. After 24 h, the protein and mRNA expression levels of DUSP26 in the chondrocytes were assessed by RT-qPCR and western blotting. Data are presented as the mean ± SD. ** P<0.01 and *** P<0.001. Representative results of three independent experiments are shown. Ad-EP, empty adenoviral vector; DUSP26, dual-specificity phosphatase 26; IL, interleukin; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin.

Article Snippet: To assess the functional integrity of chondrocytes, the P0-P1 primary chondrocytes were treated with the pro-inflammatory cytokine IL-1β (10 ng/ml; cat. no. HY-P7097; MedChemExpress) with or without the HDAC inhibitor trichostatin A (TSA; 300 nM; cat. no. T129665; Shanghai Aladdin Biochemical Technology Co., Ltd.) for 24 h at room temperature ( , ).

Techniques: Expressing, In Vitro, Isolation, Quantitative RT-PCR, Western Blot, Over Expression, Construct, Plasmid Preparation, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: DUSP26 overexpression alleviates IL-1β-induced chondrocyte injury. Rat DUSP26 overexpression adenovirus was constructed and used to infect chondrocytes. After 24 h, the chondrocytes were subjected to 10 ng/ml IL-1β. (A) Expression levels of COL2A1 and COL1A1 in the chondrocytes were detected by RT-qPCR and western blotting. (B) Relative mRNA expression levels of TNF-α and IL-6 in the chondrocytes were determined by RT-qPCR. (C) Levels of TNF-α and IL-6 in the supernatants of the chondrocytes were determined by ELISA. Two adenovirus-mediated shRNAs against rat DUSP26 were constructed and used to infect chondrocytes. After 24 h, the chondrocytes were subjected to 10 ng/ml IL-1β. (D) Expression levels of COL2A1 and COL1A1 in the chondrocytes were evaluated by RT-qPCR and western blotting. (E) Relative mRNA expression levels of TNF-α and IL-6 in the chondrocytes were determined by RT-qPCR. (F) Levels of TNF-α and IL-6 in the supernatants of the chondrocytes were detected by ELISA. Data are presented as the mean ± SD. * P<0.05, ** P<0.01 and *** P<0.001. Representative results of three independent experiments are shown. COL1A1, type I collagen; COL2A1, type II collagen; EP, empty adenoviral vector; DUSP26, dual-specificity phosphatase 26; IL, interleukin; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin.

Article Snippet: To assess the functional integrity of chondrocytes, the P0-P1 primary chondrocytes were treated with the pro-inflammatory cytokine IL-1β (10 ng/ml; cat. no. HY-P7097; MedChemExpress) with or without the HDAC inhibitor trichostatin A (TSA; 300 nM; cat. no. T129665; Shanghai Aladdin Biochemical Technology Co., Ltd.) for 24 h at room temperature ( , ).

Techniques: Over Expression, Construct, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: HDAC1, HDAC2 and HDAC8 are DUSP26-interacting proteins in chondrocytes. (A) IP/MS analysis in DUSP26-overexpressing chondrocytes under IL-1β stimulation. (B) Flow chart of target protein screening. (C) Immunohistochemical staining of HDAC1, HDAC2 and HDAC8 in the acetabular cartilage of the 4-week-old control rats and rats with DDH. (D) Protein expression levels of HDAC1, p-HDAC1, HDAC2, p-HDAC2, HDAC8 and p-HDAC8 in the chondrocytes by western blotting. (E) Co-IP experiments with DUSP26 protein from extracts of IL-1β-treated chondrocytes followed by western blotting with indicated antibodies. DEG, differentially expressed gene; DUSP26, dual-specificity phosphatase 26; HDAC, histone deacetylase; IB, immunoblot; IL, interleukin; IP, immunoprecipitation; LC-MS/MS, liquid chromatography-tandem MS; mRNA-seq, mRNA sequencing; MS, mass spectrometry; p-, phosphorylated.

Article Snippet: To assess the functional integrity of chondrocytes, the P0-P1 primary chondrocytes were treated with the pro-inflammatory cytokine IL-1β (10 ng/ml; cat. no. HY-P7097; MedChemExpress) with or without the HDAC inhibitor trichostatin A (TSA; 300 nM; cat. no. T129665; Shanghai Aladdin Biochemical Technology Co., Ltd.) for 24 h at room temperature ( , ).

Techniques: Protein-Protein interactions, Immunohistochemical staining, Staining, Control, Expressing, Western Blot, Co-Immunoprecipitation Assay, Histone Deacetylase Assay, Immunoprecipitation, Liquid Chromatography with Mass Spectroscopy, Liquid Chromatography, Sequencing, Mass Spectrometry

Journal: International Journal of Molecular Medicine

Article Title: DUSP26: Unveiling a critical molecular mediator and therapeutic target in developmental dysplasia of the hip-associated secondary osteoarthritis

doi: 10.3892/ijmm.2026.5776

Figure Lengend Snippet: Inactivation of HDAC1/2/8 inhibits DUSP26 silencing-triggered cartilage degeneration. (A) Total protein and phosphorylation levels of HDAC1, HDAC2 and HDAC8 in IL-1β-treated chondrocytes determined by western blotting. (B) Relative mRNA expression levels of HDAC1, HDAC2 and HDAC8 in chondrocytes, as determined by reverse transcription-quantitative PCR. (C) Relative mRNA expression levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after knocking down HDAC1/2/8. (D) Relative mRNA expression levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after intervention with the HDAC inhibitor TSA. (E) Protein levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after knocking down HDAC1/2/8. (F) Protein levels of COL1A1 and TNF-α in IL-1β-treated chondrocytes after intervention with the HDAC inhibitor TSA. Data are presented as the mean ± SD. * P<0.05, ** P<0.01 and *** P<0.001. (B-D) Representative results of three independent experiments are shown. COL1A1, type I collagen; DUSP26, dual-specificity phosphatase 26; HDAC, histone deacetylase; IL, interleukin; NC, negative control; ns, no significance; sh, short hairpin; TSA, trichostatin A.

Article Snippet: To assess the functional integrity of chondrocytes, the P0-P1 primary chondrocytes were treated with the pro-inflammatory cytokine IL-1β (10 ng/ml; cat. no. HY-P7097; MedChemExpress) with or without the HDAC inhibitor trichostatin A (TSA; 300 nM; cat. no. T129665; Shanghai Aladdin Biochemical Technology Co., Ltd.) for 24 h at room temperature ( , ).

Techniques: Phospho-proteomics, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Histone Deacetylase Assay, Negative Control

Journal: Molecular Medicine Reports

Article Title: Carnosic acid alleviates brain injury through NF-κB-regulated inflammation and Caspase-3-associated apoptosis in high fat-induced mouse models

doi: 10.3892/mmr.2019.10299

Figure Lengend Snippet: Primers used in the reverse transcription-quantitative PCR analysis.

Article Snippet: The primary antibodies used were as follows: IL-1β (1:100; cat. no. PA1351; Boster Biological Technology), IL-18 (1:500; cat. no. RP1017; Boster Biological Technology), TNF-α (1:500; cat. no. RP1000; Boster Biological Technology), IKKα (1:1,000; cat. no. 11930; Cell Signaling Technology, Inc.) and phosphorylated (p)-IKKα (1:1,000; cat. no. 2697; Cell Signaling Technology, Inc.), IκBα (1:500; cat. no. 4814; Cell Signaling Technology, Inc.) and p-IκBα (1:500; cat. no. 9246; Cell Signaling Technology, Inc.), NF-κB (1:1,000; cat. no. 8242; Cell Signaling Technology, Inc.) and p-NF-κB (1:1,000; cat. no. 3033; Cell Signaling Technology, Inc.), glial fibrillary acidic protein (GFAP; 1:2,000; cat. no. 3670; Cell Signaling Technology, Inc.), neuronal nuclei (Neu-N; 1:1,500; cat. no. 24307; Cell Signaling Technology, Inc.), ionized calcium-binding adapter molecule 1 (Iba-1; 1:2,000; cat. no. ab15690; Abcam), Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,500; cat. no. ab182858; Abcam), MMP-9 (1:100; cat. no. AF909; R&D Systems), Caspase-3 (1:400; cat. no. ab13585; Abcam), and GAPDH (1:1,000; cat. no. ab8245; Abcam).

Techniques:

Journal: Molecular Medicine Reports

Article Title: Carnosic acid alleviates brain injury through NF-κB-regulated inflammation and Caspase-3-associated apoptosis in high fat-induced mouse models

doi: 10.3892/mmr.2019.10299

Figure Lengend Snippet: Effects of CA on metabolic syndrome in HFD mice. (A) Body weight, (B) liver weight, (C) daily food intake and (D) daily caloric intake measured in the four experimental groups. Serum levels of (E) TG, (F) TC, (G) insulin and (H) glucose. Data are expressed as the means ± SEM. N=10/group. # P<0.05, ## P<0.01 and ### P<0.001 vs. Con; *P<0.05 and ***P<0.001 vs. Veh. CA, carnosic acid; CA-H, HFD mice treated with 20 mg/kg CA; CA-L, HFD mice treated with 10 mg/kg CA; Con, Control group; HFD, high-fat diet; TC, total cholesterol; TG, triglycerides; Veh, HFD mice.

Article Snippet: The primary antibodies used were as follows: IL-1β (1:100; cat. no. PA1351; Boster Biological Technology), IL-18 (1:500; cat. no. RP1017; Boster Biological Technology), TNF-α (1:500; cat. no. RP1000; Boster Biological Technology), IKKα (1:1,000; cat. no. 11930; Cell Signaling Technology, Inc.) and phosphorylated (p)-IKKα (1:1,000; cat. no. 2697; Cell Signaling Technology, Inc.), IκBα (1:500; cat. no. 4814; Cell Signaling Technology, Inc.) and p-IκBα (1:500; cat. no. 9246; Cell Signaling Technology, Inc.), NF-κB (1:1,000; cat. no. 8242; Cell Signaling Technology, Inc.) and p-NF-κB (1:1,000; cat. no. 3033; Cell Signaling Technology, Inc.), glial fibrillary acidic protein (GFAP; 1:2,000; cat. no. 3670; Cell Signaling Technology, Inc.), neuronal nuclei (Neu-N; 1:1,500; cat. no. 24307; Cell Signaling Technology, Inc.), ionized calcium-binding adapter molecule 1 (Iba-1; 1:2,000; cat. no. ab15690; Abcam), Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,500; cat. no. ab182858; Abcam), MMP-9 (1:100; cat. no. AF909; R&D Systems), Caspase-3 (1:400; cat. no. ab13585; Abcam), and GAPDH (1:1,000; cat. no. ab8245; Abcam).

Techniques: Control

Journal: Molecular Medicine Reports

Article Title: Carnosic acid alleviates brain injury through NF-κB-regulated inflammation and Caspase-3-associated apoptosis in high fat-induced mouse models

doi: 10.3892/mmr.2019.10299

Figure Lengend Snippet: CA decreases the inflammatory response in HFD mice. Hematoxylin and eosin staining of (A) liver and (B) pancreas indicated the pathological score in different experimental groups (magnification, ×50). Immunohistochemical analysis of (C) TNF-α and (D) IL-1β in the liver of HFD mice and percentage of positive cells. Serum levels of (E) IL-1β, (F) IL-6 and (G) TNF-α. Data are expressed as the means ± SEM. n=10 in each group. ### P<0.001 vs. Con; **P<0.01 and ***P<0.001 vs. Veh. CA, carnosic acid; CA-H, HFD mice treated with 20 mg/kg CA; CA-L, HFD mice treated with 10 mg/kg CA; Con, Control group; HFD, high-fat diet; IL, interleukin; TNF-α, tumor necrosis factor-α; Veh, HFD mice.

Article Snippet: The primary antibodies used were as follows: IL-1β (1:100; cat. no. PA1351; Boster Biological Technology), IL-18 (1:500; cat. no. RP1017; Boster Biological Technology), TNF-α (1:500; cat. no. RP1000; Boster Biological Technology), IKKα (1:1,000; cat. no. 11930; Cell Signaling Technology, Inc.) and phosphorylated (p)-IKKα (1:1,000; cat. no. 2697; Cell Signaling Technology, Inc.), IκBα (1:500; cat. no. 4814; Cell Signaling Technology, Inc.) and p-IκBα (1:500; cat. no. 9246; Cell Signaling Technology, Inc.), NF-κB (1:1,000; cat. no. 8242; Cell Signaling Technology, Inc.) and p-NF-κB (1:1,000; cat. no. 3033; Cell Signaling Technology, Inc.), glial fibrillary acidic protein (GFAP; 1:2,000; cat. no. 3670; Cell Signaling Technology, Inc.), neuronal nuclei (Neu-N; 1:1,500; cat. no. 24307; Cell Signaling Technology, Inc.), ionized calcium-binding adapter molecule 1 (Iba-1; 1:2,000; cat. no. ab15690; Abcam), Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,500; cat. no. ab182858; Abcam), MMP-9 (1:100; cat. no. AF909; R&D Systems), Caspase-3 (1:400; cat. no. ab13585; Abcam), and GAPDH (1:1,000; cat. no. ab8245; Abcam).

Techniques: Staining, Immunohistochemical staining, Control

Journal: Molecular Medicine Reports

Article Title: Carnosic acid alleviates brain injury through NF-κB-regulated inflammation and Caspase-3-associated apoptosis in high fat-induced mouse models

doi: 10.3892/mmr.2019.10299

Figure Lengend Snippet: Effects of CA on the secretion of pro-inflammatory cytokines in the brain of HFD mice. (A) Immunohistochemical analysis of IL-1β and TNF-α in brain tissues of HFD mice (magnification, ×50). (B) Protein expression levels of IL-1β, IL-18 and TNF-α in HFD mice, as assessed by western blot analysis. (C) mRNA expression levels of pro-inflammatory cytokines in the brain of HFD mice, as assessed by reverse transcription-quantitative PCR analysis. Data are expressed as the means ± SEM. n=10 in each group. ### P<0.001 vs. Con; *P<0.05, **P<0.01 and ***P<0.001 vs. Veh. CA, carnosic acid; CA-H, HFD mice treated with 20 mg/kg CA; CA-L, HFD mice treated with 10 mg/kg CA; Con, Control group; HFD, high-fat diet; IL, interleukin; TNF-α, tumor necrosis factor-α; Veh, HFD mice.

Article Snippet: The primary antibodies used were as follows: IL-1β (1:100; cat. no. PA1351; Boster Biological Technology), IL-18 (1:500; cat. no. RP1017; Boster Biological Technology), TNF-α (1:500; cat. no. RP1000; Boster Biological Technology), IKKα (1:1,000; cat. no. 11930; Cell Signaling Technology, Inc.) and phosphorylated (p)-IKKα (1:1,000; cat. no. 2697; Cell Signaling Technology, Inc.), IκBα (1:500; cat. no. 4814; Cell Signaling Technology, Inc.) and p-IκBα (1:500; cat. no. 9246; Cell Signaling Technology, Inc.), NF-κB (1:1,000; cat. no. 8242; Cell Signaling Technology, Inc.) and p-NF-κB (1:1,000; cat. no. 3033; Cell Signaling Technology, Inc.), glial fibrillary acidic protein (GFAP; 1:2,000; cat. no. 3670; Cell Signaling Technology, Inc.), neuronal nuclei (Neu-N; 1:1,500; cat. no. 24307; Cell Signaling Technology, Inc.), ionized calcium-binding adapter molecule 1 (Iba-1; 1:2,000; cat. no. ab15690; Abcam), Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,500; cat. no. ab182858; Abcam), MMP-9 (1:100; cat. no. AF909; R&D Systems), Caspase-3 (1:400; cat. no. ab13585; Abcam), and GAPDH (1:1,000; cat. no. ab8245; Abcam).

Techniques: Immunohistochemical staining, Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Control

Journal: Molecular Medicine Reports

Article Title: Carnosic acid alleviates brain injury through NF-κB-regulated inflammation and Caspase-3-associated apoptosis in high fat-induced mouse models

doi: 10.3892/mmr.2019.10299

Figure Lengend Snippet: CA attenuates brain injury in HFD mice by inactivating the NF-κB signaling pathway. (A) Protein expression levels of p-IKKα, IKKα, GFAP and GAPDH in different groups, as assessed by western blot analysis. (B) Protein expression levels of p-IκBα, IκBα, Neu-N and GAPDH in different groups, as assessed by western blot analysis. (C) Protein expression levels of p-NF-κB, NF-κB, Iba-1 and GAPDH in different groups, as assessed by western blot analysis. Data are expressed as the means ± SEM. n=10 in each group. ## P<0.01 and ### P<0.001 vs. Con; *P<0.05, **P<0.01 and ***P<0.001 vs. Veh. CA, carnosic acid; CA-H, HFD mice treated with 20 mg/kg CA; CA-L, HFD mice treated with 10 mg/kg CA; Con, Control group; GFAP, glial fibrillary acidic protein; HFD, high-fat diet; IκBα, NF-κB inhibitor α; IKKα, IκB kinase α; Neu-N, neuronal nuclei; Iba-1, ionized calcium-binding adapter molecule 1; p, phosphorylated; Veh, HFD mice.

Article Snippet: The primary antibodies used were as follows: IL-1β (1:100; cat. no. PA1351; Boster Biological Technology), IL-18 (1:500; cat. no. RP1017; Boster Biological Technology), TNF-α (1:500; cat. no. RP1000; Boster Biological Technology), IKKα (1:1,000; cat. no. 11930; Cell Signaling Technology, Inc.) and phosphorylated (p)-IKKα (1:1,000; cat. no. 2697; Cell Signaling Technology, Inc.), IκBα (1:500; cat. no. 4814; Cell Signaling Technology, Inc.) and p-IκBα (1:500; cat. no. 9246; Cell Signaling Technology, Inc.), NF-κB (1:1,000; cat. no. 8242; Cell Signaling Technology, Inc.) and p-NF-κB (1:1,000; cat. no. 3033; Cell Signaling Technology, Inc.), glial fibrillary acidic protein (GFAP; 1:2,000; cat. no. 3670; Cell Signaling Technology, Inc.), neuronal nuclei (Neu-N; 1:1,500; cat. no. 24307; Cell Signaling Technology, Inc.), ionized calcium-binding adapter molecule 1 (Iba-1; 1:2,000; cat. no. ab15690; Abcam), Bax (1:1,000; cat. no. ab32503; Abcam), Bcl-2 (1:1,500; cat. no. ab182858; Abcam), MMP-9 (1:100; cat. no. AF909; R&D Systems), Caspase-3 (1:400; cat. no. ab13585; Abcam), and GAPDH (1:1,000; cat. no. ab8245; Abcam).

Techniques: Expressing, Western Blot, Control, Binding Assay

Journal: PloS one

Article Title: Deficiency of Smad7 enhances cardiac remodeling induced by angiotensin II infusion in a mouse model of hypertension.

doi: 10.1371/journal.pone.0070195

Figure Lengend Snippet: Figure 4. Loss of Smad7 aggravates proinflammation expression during cardiac inflammation in response to Ang II infusion. (A) IL- 1b expression. (B) TNF-a expression. (i) Immunohistochemistry (IHC); (ii) quantitative analysis of immunohistochemical staining (IHC); (iii) real-time PCR analysis. Data represent the mean6SE for a group of 6 mice. *P,0.05, ***P,0.001 vs saline control; #P,0.05, ##P,0.01, ###P,0.001 vs Ang II– infused Smad7 WT mice. Scale bar = 100 mm. doi:10.1371/journal.pone.0070195.g004

Article Snippet: The antibodies used in this study included: collagen I (Southern Biotech Inc., PLOS ONE | www.plosone.org 3 July 2013 | Volume 8 | Issue 7 | e70195 Birmingham, AL), a-SMA(R&D, Minneapolis, MN), TNFa and IL-1b (Santa Cruz Biotechnology, Santa Cruz, CA), CD3+T cells (Abcam, Cambridge, UK), macrophages (F4/80+) (Serotec, Oxford, UK).

Techniques: Expressing, Immunohistochemistry, Immunohistochemical staining, Staining, Real-time Polymerase Chain Reaction, Saline, Control