Journal:

Article Title: Human Cytomegalovirus UL99-Encoded pp28 Is Required for the Cytoplasmic Envelopment of Tegument-Associated Capsids

doi: 10.1128/JVI.77.19.10594-10605.2003

Figure Lengend Snippet: Accumulation of viral proteins after infection with wild-type (wt) and pp28-deficient viruses. Normal fibroblasts were mock-infected (M) or infected with BADwt, BADsubUL99, or BADpmUL99 at a multiplicity of infection of 0.01 PFU/cell, and whole-cell extracts were prepared at 72 h (A) or at the indicated times after infection (days postinfection [dpi]) (B). Proteins were analyzed by Western blot analysis with antibodies specific for the UL123-encoded IE1, pUL97, UL83-encoded pp65, UL82-encoded pp71, pUL44, or UL99-encoded pp28.

Article Snippet: Primary antibodies used in these studies were UL32-encoded pp150 and UL83-encoded pp65 rabbit polyclonal antibodies (gift of M. Schrader, University of Marburg), UL99-encoded pp28 mouse monoclonal antibody (10B4-29), UL55-encoded gB (58-15 monoclonal antibody produced by immunization with disrupted virions) (P. Robinson and T. Shenk, unpublished data).

Techniques: Infection, Western Blot

Journal:

Article Title: Human Cytomegalovirus UL99-Encoded pp28 Is Required for the Cytoplasmic Envelopment of Tegument-Associated Capsids

doi: 10.1128/JVI.77.19.10594-10605.2003

Figure Lengend Snippet: Localization of viral proteins expressed in normal fibroblasts infected with wild-type (WT) and pp28-deficient viruses. Cells were infected with BADwt or BADsubUL99 at a multiplicity of infection of 0.01 PFU/cell and processed for immunofluorescence at 120 h postinfection. Four images are presented for each cell. The upper-left quadrant shows UL32-encoded pp150 (A), UL83-encoded pp65 (B), or UL55-encoded gB (C) in red. The lower-left quadrants of panels A and B display pp28 in blue, whereas the lower-left panel of C contains blue-stained nuclei. In all panels, the upper-right quadrant displays Golgi in green and the lower-right panel shows a multicolored merged image.

Article Snippet: Primary antibodies used in these studies were UL32-encoded pp150 and UL83-encoded pp65 rabbit polyclonal antibodies (gift of M. Schrader, University of Marburg), UL99-encoded pp28 mouse monoclonal antibody (10B4-29), UL55-encoded gB (58-15 monoclonal antibody produced by immunization with disrupted virions) (P. Robinson and T. Shenk, unpublished data).

Techniques: Infection, Immunofluorescence, Staining

Journal: The Kaohsiung journal of medical sciences

Article Title: Investigating the Impact of hsa_circ_0005255 on Proliferation and Autophagy in Crohn's Disease Intestinal Epithelial Cells Through miR-23a-3p-Mediated NCOA3 Expression.

doi: 10.1002/kjm2.70035

Figure Lengend Snippet: FIGURE 2 | Overexpression of hsa_circ_0005255 significantly improves intestinal epithelial cell proliferation and autophagy in CD mice. CD mouse model was induced using DSS and AAV overexpression vector targeting hsa_circ_0005255 was injected into CD mice. (A) RT-qPCR con- firmed that hsa_circ_0005255 was lowly expressed in CD mice and was reversed by AAV-oe-hsa_circ_0005255; (B) DAI scores of each group of mice; (C) Representative images of HE staining of colonic tissues of mice in each group (Yellow arrows indicate inflammatory cell infiltration); (D) TUNEL staining to detect apoptosis in colonic tissues of mice in each group (green fluorescence to indicate TUNE positive cells and blue fluores- cence for DAPI to label nuclei); (E) ELISA to detect the levels of inflammatory factors TNF-α, IL-1β, and IL-6 in the serum of mice in each group; (F) Immunofluorescence to detect the expression of ZO-1 and F4/80 in the colonic tissues of mice in each group (F4/80 is a macrophage marker. zo- 1 is a tight junction protein marker); (G) ELISA to detect the serum levels of LPS; (H) Western blot experiments to detect the expression of cleaved caspase-3, p-p65, LC3B, p62, and Beclin-1 in colon tissues of mice in each group; data are expressed as mean ± SD (N = 8).

Article Snippet: The primary antibodies employed were cleaved caspase- 3 (ab2302, Abcam), phospho- p65 (p- p65, 3031, Cell Signaling Technology), p65 (6956, Cell Signaling Technology), LC3B (2775, Cell Signaling Technology), p62 (5114, Cell Signaling Technology), and Beclin- 1 (3495, Cell Signaling Technology).

Techniques: Over Expression, Plasmid Preparation, Injection, Quantitative RT-PCR, Staining, TUNEL Assay, Fluorescence, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Expressing, Marker, Western Blot

Journal: The Kaohsiung journal of medical sciences

Article Title: Investigating the Impact of hsa_circ_0005255 on Proliferation and Autophagy in Crohn's Disease Intestinal Epithelial Cells Through miR-23a-3p-Mediated NCOA3 Expression.

doi: 10.1002/kjm2.70035

Figure Lengend Snippet: FIGURE 3 | Overexpression of hsa_circ_0005255 ameliorates LPS-induced intestinal epithelial cell injury in vitro. Caco2 cells were treated with LPS to establish an in vitro model of CD, and pcDNA 3.1-hsa_circ_0005255 was transfected into Caco2 cells. (A) RT-qPCR to detect the expression of hsa_circ_0005255; (B): CCK-8 assay to detect the proliferation rate; (C) TEER test to assess the tight junction function of cells; (D) Flow cytometry to detect the apoptosis rate of cells; (E) ELISA kits to detect the levels of inflammatory factors TNF-α, IL-1β and IL-6 in cells; (F) MDC staining to detect the formation of autophagosomes in cells; (G) Western blot assay to detect the expression of ZO-1, Ki-67, cleaved caspase-3, p-p65, LC3B, p62, and Beclin-1 expression in cells; data are expressed as mean ± SD (N = 3).

Article Snippet: The primary antibodies employed were cleaved caspase- 3 (ab2302, Abcam), phospho- p65 (p- p65, 3031, Cell Signaling Technology), p65 (6956, Cell Signaling Technology), LC3B (2775, Cell Signaling Technology), p62 (5114, Cell Signaling Technology), and Beclin- 1 (3495, Cell Signaling Technology).

Techniques: Over Expression, In Vitro, Transfection, Quantitative RT-PCR, Expressing, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Western Blot

Journal: The Kaohsiung journal of medical sciences

Article Title: Investigating the Impact of hsa_circ_0005255 on Proliferation and Autophagy in Crohn's Disease Intestinal Epithelial Cells Through miR-23a-3p-Mediated NCOA3 Expression.

doi: 10.1002/kjm2.70035

Figure Lengend Snippet: FIGURE 5 | hsa_circ_0005255 ameliorates LPS-induced intestinal epithelial cell injury in vitro by modulating miR-23a-3p. Caco2 cells were treated with LPS to establish an in vitro model of CD, and pcDNA 3.1-hsa_circ_0005255 and miR-23a-3p mimic were co-transfected into Caco2 cells. (A) RT-qPCR to detect the expression of miR-23a-3p in cells; (B) CCK-8 assay to detect the proliferation rate of cells; (C) TEER test to assess the tight junction function of cells; (D) Flow cytometry to detect the apoptosis rate of cells; (E) ELISA kits to assess the levels of inflammatory factors TNF-α, IL-1β and IL-6 of cells; (F) MDC staining to detect the formation of autophagosomes of cells (green fluorescence indicates autophagic vesicles, blue fluorescence indicates nuclei); (G) Western blot assay to detect the expression of ZO-1, Ki-67, cleaved caspase-3, p-p65, LC3B, p62 and Beclin-1 ex- pression in cells; data are expressed as mean ± SD (N = 3).

Article Snippet: The primary antibodies employed were cleaved caspase- 3 (ab2302, Abcam), phospho- p65 (p- p65, 3031, Cell Signaling Technology), p65 (6956, Cell Signaling Technology), LC3B (2775, Cell Signaling Technology), p62 (5114, Cell Signaling Technology), and Beclin- 1 (3495, Cell Signaling Technology).

Techniques: In Vitro, Transfection, Quantitative RT-PCR, Expressing, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Fluorescence, Western Blot

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: Journal of Food Biochemistry

Article Title: Betulinic Acid–Mediated NR1D1 Alleviates Dextran Sulfate Sodium–Induced Colitis in Mice by Modulating VDAC1/NF‐κВ

doi: 10.1155/jfbc/3244032

Figure Lengend Snippet: Figure 4: BHA alleviates infammatory response in DSS-induced colitis. (a) Te gene expression of IL-1β, IL-6, TNF-α, and IL-18. (b) Te content of IL-1β and IL-18 in colon. (c) NR1D1, VDAC1, p65, and pp65 gray values. (d) Te protein of NR1D1 and VDAC1 and the ratio of pp65 to p65. Data were analyzed by using one-way ANOVA followed by the Duncan’s test. Te signifcance indicated refers to the comparison between controls and all treatment groups.

Article Snippet: Te primary antibodies applied included NR1D1 (WH0009572M2, Sigma-Aldrich, MO), VDAC1 (ab306581, Abcam, Cambridge, UK), pp65 (3033, CST, China), p65 (8242, CST, China), and β-actin (ab8226, Abcam, Cambridge, UK), following the manufacturers’ instructions.

Techniques: Gene Expression, Comparison

Journal: Journal of Food Biochemistry

Article Title: Betulinic Acid–Mediated NR1D1 Alleviates Dextran Sulfate Sodium–Induced Colitis in Mice by Modulating VDAC1/NF‐κВ

doi: 10.1155/jfbc/3244032

Figure Lengend Snippet: Figure 7: Efect of BHA on infammatory response in LPS-induced CCD841. (a) Te gene expression of IL-1β, IL-6, TNF-α and IL-18. (b) Te content of IL-1β and IL-18. (c) NR1D1, VDAC1, p65, and pp65 gray values. (d) Te protein of NR1D1 and VDAC1 and the ratio of pp65 to p65. (e) Immunofuorescence of NR1D1. Data were analyzed by using one-way ANOVA followed by the Duncan’s test. Te signifcance indicated refers to the comparison between controls and all treatment groups.

Article Snippet: Te primary antibodies applied included NR1D1 (WH0009572M2, Sigma-Aldrich, MO), VDAC1 (ab306581, Abcam, Cambridge, UK), pp65 (3033, CST, China), p65 (8242, CST, China), and β-actin (ab8226, Abcam, Cambridge, UK), following the manufacturers’ instructions.

Techniques: Gene Expression, Comparison

Journal: Journal of Food Biochemistry

Article Title: Betulinic Acid–Mediated NR1D1 Alleviates Dextran Sulfate Sodium–Induced Colitis in Mice by Modulating VDAC1/NF‐κВ

doi: 10.1155/jfbc/3244032

Figure Lengend Snippet: Figure 9: Efect of NR1D1 on genes and proteins related to infammation. Generation of NR1D1 knockdown cell using siRNA. (a) NR1D1 gray values. (b) NR1D1 protein expression levels. (c) Te genes expression of IL-1β, IL-6, TNF-α and IL-18. (d) VDAC1, p65, and pp65 gray values. (e) Te protein of VDAC1 and the ratio of pp65 to p65. Data were analyzed by using one-way ANOVA followed by the Duncan’s test. Te signifcance indicated refers to the comparison between controls and all treatment groups.

Article Snippet: Te primary antibodies applied included NR1D1 (WH0009572M2, Sigma-Aldrich, MO), VDAC1 (ab306581, Abcam, Cambridge, UK), pp65 (3033, CST, China), p65 (8242, CST, China), and β-actin (ab8226, Abcam, Cambridge, UK), following the manufacturers’ instructions.

Techniques: Knockdown, Expressing, Comparison

Journal: Experimental and Therapeutic Medicine

Article Title: LncRNA NEAT1 promotes apoptosis and inflammation in LPS-induced sepsis models by targeting miR-590-3p

doi: 10.3892/etm.2020.9079

Figure Lengend Snippet: Inhibition of NEAT1 suppresses the NF-κB signaling pathway via targeting miR-590-3p. H9c2 cells were transfected with si-NEAT1 and anti-miR-590-3p or their corresponding negative controls (si-NC and anti-NC), followed by treatment with LPS. (A) The protein levels of NF-κB pathway-related factors p-p65 and p65 were detected and (B) quantified using western blot analysis. * P<0.05. NEAT1, nuclear enriched abundant transcript 1; miR, microRNA; si-NEAT1, small interfering RNA targeting NEAT1; LPS, lipopolysaccharide; p, phosphorylated.

Article Snippet: Primary antibodies against Bcl-2 (1:500; cat. no. BA0412), Bax (1:1,000; cat. no. BA0315-2), caspase 3 (1:1,000; cat. no. BA3257), TNF receptor associated factor 6 (TRAF6; 1:1,500; cat. no. A00185), phosphorylated (p)-p65 (1:1,000; cat. no. P00284), p65 (1:2,000; cat. no. A00284) and GAPDH (1:2,000; cat. no. BA2913) were purchased from Wuhan Boster Biological Technology, Ltd. Membranes were then incubated with horseradish peroxidase-labeled secondary antibody (1:5,000; cat no. BA1056; Wuhan Boster Biological Technology, Ltd.) for 1 h at room temperature.

Techniques: Inhibition, Transfection, Western Blot, Small Interfering RNA