Journal: International journal of molecular sciences

Article Title: WGA-M001, a Mixture of Total Extracts of Tagetes erecta and Ocimum basilicum , Synergistically Alleviates Cartilage Destruction by Inhibiting ERK and NF-κB Signaling.

doi: 10.3390/ijms242417459

Figure Lengend Snippet: Figure 4. WGA-M001 regulated OA-related molecules through dephosphorylation in the NF-κB and ERK pathways. For in silico analysis, 200 µg/mL of Tagetes erecta, Ocimum basilicum, or WGA-M001 was used for treatment of chondrocytes for 12 h along with IL-1β (1 ng/mL), and RNA sequencing was performed. (A) In each signaling pathway, the number of genes upregulated by IL-1β and then downregulated by T. erecta, O. basilicum, or WGA-M001 was shown. (B,C) Protein levels of pp38, p38, pJNK, JNK, pERK, ERK, pp65, p65, and IκB were detected by Western blot analysis and relative intensities were quantified by densitometry (n = 5). Each protein level was normalized to ERK. Data are represented as mean ± SD as results of analysis by one-way ANOVA with Dunnett’s multiple comparisons test (n = 5). * p < 0.05, **** p < 0.0001, and n.s = not significant.

Article Snippet: 2023, 24, 17459 12 of 16 cam), rabbit anti-COX-2 (ab52237; Abcam), mouse anti-IκB (9242; Cell Signaling Technology (CST), Danvers, MA, USA), mouse anti-p65 (#6956; CST), mouse anti-pp65 (#13346; CST), mouse anti-p38 (#9212; CST), mouse anti-pp38 (#9215S; CST), mouse anti-c-JNK (#9252S; CST), mouse anti-pJNK (#9251S; CST), and mouse anti-pERK (#9101S; CST).

Techniques: De-Phosphorylation Assay, In Silico, RNA Sequencing, Western Blot

Journal: Mediators of Inflammation

Article Title: MCL Plays an Anti-Inflammatory Role in Mycobacterium tuberculosis -Induced Immune Response by Inhibiting NF- κ B and NLRP3 Inflammasome Activation

doi: 10.1155/2017/2432904

Figure Lengend Snippet: MCL inhibits Mtb-triggered NF- κ B activation. Raw264.7 cells (1 × 10 6 /well) were seeded in 6-well plates and infected with Mtb for 4 hrs. Cells were washed by PBS and treated with MCL (10 μ M) for indicated time courses. Phosphorylation of p65 and β -actin was detected by Western blot (a). The lower bar graph showed the statistical results for phosphorylation levels of NF- κ B p65 (a). Data are presented as mean ± SD in at least three independent experiments, ∗∗ p < 0.01. Raw264.7 cells (3 × 10 5 /well) were cotransfected with NF- κ B luciferase reporter plasmid and pRL-TKR Renilla luciferase plasmid as previous study . After 30 hr, cells were infected with H37Ra at MOI of 10 and treated with MCL (10 μ M) for 12 hr, and the luciferase activities were measured (b). Data are shown with the means ± SD ( n ≧3). ∗ p < 0.05. Raw264.7cells (3 × 10 5 /well) were plated in climbing slice and followed by appropriate treatment. Immunofluorescence assay was conducted with rabbit anti-pp65 antibody (green) and DAPI (blue) (c). The results represented three independent experiments.

Article Snippet: Rabbit anti-ASC (10500-1-AP, proteintech), anti-LC3 (sc-16755, Santa Cruz Biotechnology), anti-caspase-1 (sc-514, Santa Cruz Biotechnology), and anti-pp65 (cst-3033, cell signaling technology) antibodies were used for immunoblot.

Techniques: Activation Assay, Infection, Western Blot, Luciferase, Plasmid Preparation, Immunofluorescence

Journal: Journal of Inflammation Research

Article Title: PM2.5 Promotes Macrophage-Mediated Inflammatory Response Through Airway Epithelial Cell-Derived Exosomal miR-155-5p

doi: 10.2147/jir.s482509

Figure Lengend Snippet: Figure 1 PM2.5-BEAS-2B co-cultured with Mφ. (A) PM2.5-BEAS-2B cell co-cultured with Mφ mode diagram; (B) Cellular supernatant IL-6, IL-1β, TNF-α contents of Mφ co-cultured with PM2.5-BEAS-2B cells; (C) mRNA expression levels of IL-6, IL-1β, TNF-α in Mφ co-cultured with PM2.5-BEAS-2B cells; (D) miR-155-5p expression level in Mφ co-cultured with PM2.5-BEAS-2B cells; (E) mRNA expression levels of SOCS1, NF-κB in Mφ co-cultured with PM2.5-BEAS-2B cells; (F and G) Protein expression level of SOCS1, P65, PP65 in Mφ co-cultured with PM2.5-BEAS-2B cells as detected by Western blot analysis as well as statistical analysis of gray values. Experimental group: group A PM2.50-BEAS-2B+Mφ, group B PM2.5100-BEAS-2B+Mφ, group C PM2.5200-BEAS-2B+Mφ, group D PM2.5400-BEAS-2B+Mφ. **P<0.01, *P<0.05 vs group A.

Article Snippet: After blocking with protein-free rapid-blocking solution (Boster, Wuhan, China) for 1h, the membranes were exposed to rabbit anti-SOCS1 polyclonal antibody (Boster), rabbit anti-P65 polyclonal antibody (Boster), rabbit anti-PP65 monoclonal antibody (CellSignalingTechnology), rabbit anti-βtublin polyclonal antibody (Boster), and rabbit anti-GAPDH polyclonal antibody (Boster) at 4°C overnight.

Techniques: Cell Culture, Expressing, Western Blot

Journal: Journal of Inflammation Research

Article Title: PM2.5 Promotes Macrophage-Mediated Inflammatory Response Through Airway Epithelial Cell-Derived Exosomal miR-155-5p

doi: 10.2147/jir.s482509

Figure Lengend Snippet: Figure 4 PM2.5-BEAS-2B-Exo co-cultured with Mφ. (A) PM2.5-BEAS-2B-Exo co-cultured with Mφ mode diagram; (B) Cellular supernatant IL-6, IL-1β, TNF-α contents of Mφ incubated with PM2.5-BEAS-2B-Exo; (C) mRNA expression levels of IL-6, IL-1β, TNF-α in Mφ incubated with PM2.5-BEAS-2B-Exo; (D) miR-155-5p expression level in Mφ incubated with PM2.5-BEAS-2B-Exo; (E) mRNA expression levels of SOCS1, NF-κB in Mφ incubated with PM2.5-BEAS-2B-Exo; (F and G) Protein expression level of SOCS1, P65, PP65 in Mφ incubated with PM2.5-BEAS-2B-Exo as detected by Western blot analysis as well as statistical analysis of gray values. Experimental group: group E PM2.50-BEAS-2B-Exo+Mφ, group F PM2.5100-BEAS-2B-Exo+Mφ, group G PM2.5200-BEAS-2B-Exo+Mφ, group H PM2.5400-BEAS-2B-Exo+Mφ. **P<0.01, *P<0.05 vs group E.

Article Snippet: After blocking with protein-free rapid-blocking solution (Boster, Wuhan, China) for 1h, the membranes were exposed to rabbit anti-SOCS1 polyclonal antibody (Boster), rabbit anti-P65 polyclonal antibody (Boster), rabbit anti-PP65 monoclonal antibody (CellSignalingTechnology), rabbit anti-βtublin polyclonal antibody (Boster), and rabbit anti-GAPDH polyclonal antibody (Boster) at 4°C overnight.

Techniques: Cell Culture, Incubation, Expressing, Western Blot

Journal: Journal of Inflammation Research

Article Title: PM2.5 Promotes Macrophage-Mediated Inflammatory Response Through Airway Epithelial Cell-Derived Exosomal miR-155-5p

doi: 10.2147/jir.s482509

Figure Lengend Snippet: Figure 5 GW4869 improves inflammation in Mφ. PM2.5-BEAS-2B cells were pretreated with GW4869 and co-cultured with Mφ. (A) Cellular supernatant IL-6, IL-1β, TNF-α contents of Mφ; (B) mRNA expression levels of IL-6, IL-1β, TNF-α in Mφ; (C) miR-155-5p expression level in Mφ; (D) mRNA expression levels of SOCS1, NF-κB in Mφ; (E and F) Protein expression level of SOCS1, P65, PP65 in Mφ as detected by Western blot analysis as well as statistical analysis of gray values. Experimental group: group A PM2.50-BEAS-2B+Mφ, group C PM2.5200-BEAS-2B+Mφ, group I PM2.5200-BEAS-2B+GW4869+Mφ. **P<0.01, *P<0.05 vs group A;▲▲P<0.01,▲P<0.05 vs group C.

Article Snippet: After blocking with protein-free rapid-blocking solution (Boster, Wuhan, China) for 1h, the membranes were exposed to rabbit anti-SOCS1 polyclonal antibody (Boster), rabbit anti-P65 polyclonal antibody (Boster), rabbit anti-PP65 monoclonal antibody (CellSignalingTechnology), rabbit anti-βtublin polyclonal antibody (Boster), and rabbit anti-GAPDH polyclonal antibody (Boster) at 4°C overnight.

Techniques: Cell Culture, Expressing, Western Blot

Journal: Virology Journal

Article Title: In vitro study on human cytomegalovirus affecting early pregnancy villous EVT's invasion function

doi: 10.1186/1743-422X-8-114

Figure Lengend Snippet: Histology of two groups'villous explants under HE staining . A. Human villous explants in vitro HE ×200; B. Human villous explants infected by HCMV in vitro HE ×200.

Article Snippet: DMEM/Ham's F 12 culture medium (Gibco), standard fetal bovine serum (Hyclone), rabbit anti HCMV pp65 polyclonal antibody (Santa), rabbit anti c-erbB-2 polyclonal antibody, rabbit anti MMP-2 and MMP-9 polyclonal antibodies (Wuhan Boster Bio-engineering Co., Ltd), SP immunohistochemistry kit (Zhongshan Goldenbridge Biotechnology Co., Ltd), total protein extraction kit (BestBio), β-actin loading control antibody (Beijing Biosynthesis Biotechnology Co., Ltd) and so on.

Techniques: Staining, In Vitro, Infection

Journal: Virology Journal

Article Title: In vitro study on human cytomegalovirus affecting early pregnancy villous EVT's invasion function

doi: 10.1186/1743-422X-8-114

Figure Lengend Snippet: The expression of HCMVpp65 antigen in two groups'villous explants detected by immunohistostaining . A. The expression of HCMVpp65 antigen in normal human villous explants detected by immunohistostaining ×400; B. The expression of HCMVpp65 antigen in human villous explants infected by HCMV detected by immunohistostaining ×400.

Article Snippet: DMEM/Ham's F 12 culture medium (Gibco), standard fetal bovine serum (Hyclone), rabbit anti HCMV pp65 polyclonal antibody (Santa), rabbit anti c-erbB-2 polyclonal antibody, rabbit anti MMP-2 and MMP-9 polyclonal antibodies (Wuhan Boster Bio-engineering Co., Ltd), SP immunohistochemistry kit (Zhongshan Goldenbridge Biotechnology Co., Ltd), total protein extraction kit (BestBio), β-actin loading control antibody (Beijing Biosynthesis Biotechnology Co., Ltd) and so on.

Techniques: Expressing, Infection

Journal: Virology Journal

Article Title: In vitro study on human cytomegalovirus affecting early pregnancy villous EVT's invasion function

doi: 10.1186/1743-422x-8-114

Figure Lengend Snippet: Figure 1 Histology of two groups’villous explants under HE staining. A. Human villous explants in vitro HE ×200; B. Human villous explants infected by HCMV in vitro HE ×200.

Article Snippet: DMEM/Ham’s F 12 culture medium (Gibco), standard fetal bovine serum (Hyclone), rabbit anti HCMV pp65 polyclonal antibody (Santa), rabbit anti c-erbB-2 polyclonal antibody, rabbit anti MMP-2 and MMP-9 polyclonal antibodies (Wuhan Boster Bio-engineering Co., Ltd), SP immunohistochemistry kit (Zhongshan Goldenbridge Biotechnology Co., Ltd), total protein extraction kit (BestBio), b-actin loading control antibody (Beijing Biosynthesis Biotechnology Co., Ltd) and so on.

Techniques: Staining, In Vitro, Infection

Journal: Virology Journal

Article Title: In vitro study on human cytomegalovirus affecting early pregnancy villous EVT's invasion function

doi: 10.1186/1743-422x-8-114

Figure Lengend Snippet: Figure 2 The expression of HCMVpp65 antigen in two groups’villous explants detected by immunohistostaining. A. The expression of HCMVpp65 antigen in normal human villous explants detected by immunohistostaining ×400; B. The expression of HCMVpp65 antigen in human villous explants infected by HCMV detected by immunohistostaining ×400.

Article Snippet: DMEM/Ham’s F 12 culture medium (Gibco), standard fetal bovine serum (Hyclone), rabbit anti HCMV pp65 polyclonal antibody (Santa), rabbit anti c-erbB-2 polyclonal antibody, rabbit anti MMP-2 and MMP-9 polyclonal antibodies (Wuhan Boster Bio-engineering Co., Ltd), SP immunohistochemistry kit (Zhongshan Goldenbridge Biotechnology Co., Ltd), total protein extraction kit (BestBio), b-actin loading control antibody (Beijing Biosynthesis Biotechnology Co., Ltd) and so on.

Techniques: Expressing, Infection