Journal: International journal of immunopathology and pharmacology

Article Title: Corilagin relieves atherosclerosis via the toll-like receptor 4 signaling pathway in vascular smooth muscle cells.

doi: 10.1177/03946320241254083

Figure Lengend Snippet: Figure 2. Effect of ox-LDL on the TLR4 signaling pathway in MOVAS cells. (a) mRNA expression of TLR4, TIRAP and MyD88 was measured by qRT-PCR. *p < .05 compared with other concentrations of ox-LDL groups or control group as determined by one-way ANOVA and subsequent Student–Newman–Keuls q-test (n = 5). (b) *p < .05 compared with either time point or control groups determined by one-way ANOVA and subsequent Student–Newman–Keuls q-test (n = 5).

Article Snippet: Afterwards, the cells were subjected to a 2-h incubation at 37°C with either primary anti-TLR4 antibody (1:200 dilution; Cat No. BA1717; Boster Biological Technology, Pleasanton, CA, USA) or anti-MyD88 antibody (1:400 dilution; Cat No. PB9148; Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: International journal of immunopathology and pharmacology

Article Title: Corilagin relieves atherosclerosis via the toll-like receptor 4 signaling pathway in vascular smooth muscle cells.

doi: 10.1177/03946320241254083

Figure Lengend Snippet: Figure 3. Effect of corilagin on the TLR4 signaling pathway in MOVAS cells stimulated by ox-LDL. (a) mRNA expression of TLR4, TIRAP, MyD88, TRAF6, p38, NEMO and IRF5 was measured by qRT-PCR. *p < .05 compared with the control group, #p < .05 compared with the ox-LDL group, **p < .05 compared with the aspirin group as determined by one-way ANOVA and subsequent Student–Newman–Keuls q-test (n = 5). (B. C) Protein abundance was measured by western blotting. *p < .05 compared with the control group, #p < .05 compared with the ox-LDL group, **p < .05 compared with the aspirin group, as determined by one-way ANOVA and subsequent Student–Newman–Keuls q-test (n = 5). (D, E) Abundance of IL-6 and MCP-1 in cell culture supernatant was measured by ELISAs. *p < .05 compared with the control group, #p < .05 compared with the ox-LDL group, **p < .05 compared with the aspirin group as determined by one-way ANOVA and subsequent Student–Newman–Keuls q-test (n = 5). (F) Effect of corilagin on the MOVAS cells proliferation. *p < .05 compared with the control group, #p < .05 compared with the ox-LDL group determined by one-way ANOVA and subsequent Student–Newman–Keuls q-test (n = 5). (G, H) apoptosis ratio of MOVAS were detected by Annexin V staining. No significant difference was found among four groups determined by one-way ANOVA and subsequent Student–Newman–Keuls q-test (n = 5).

Article Snippet: Afterwards, the cells were subjected to a 2-h incubation at 37°C with either primary anti-TLR4 antibody (1:200 dilution; Cat No. BA1717; Boster Biological Technology, Pleasanton, CA, USA) or anti-MyD88 antibody (1:400 dilution; Cat No. PB9148; Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Expressing, Quantitative RT-PCR, Control, Quantitative Proteomics, Western Blot, Cell Culture, Staining

Journal:

Article Title: Expression of RASSF1A, an epigenetically silenced tumor suppressor, overcomes resistance to apoptosis induction by interferons

doi: 10.1158/0008-5472.CAN-05-2303

Figure Lengend Snippet: A): Map of lentiviral RASSF1A construct. B): ACHN cells were transduced with RASSF1A carrying lentivirus or empty virus. 72h later selective antibiotic was added and the population of clones was grown in selection media for 14 days before assessing apoptotic response to IFN-β (50 U/ml over 5 days) by TUNEL assay. After confirmation of stable expression of RASSF1A by immunoblotting, clonal selection was performed to identify clones that expressed similar amounts of RASSF1A protein as achieved by DNMT1 AS (40 nM) over 8 days. Treatment with DNMT1 mismatch control oligonucleotide (MM) and empty lentivirus transduced cells served as negative controls. In clones expressing RASSF1A at levels comparable to DNA demethylating treatment, IFN-β (50 U/ml over 5 days) resulted in 20–40% apoptotic cells while stable transduction with empty lentivirus (kept in selective antibiotic for equal amounts of time) did not. C): Concurrent treatment of ACHN RASSF1A clone 1.3 with 2 μg/ml TRAIL neutralizing antibody (TRAIL-N AB) and IFN-β (50 U/ml) over 5 days inhibited IFN-induced apoptosis compared to cotreatment with 2 μg/ml control rabbit immunoglobulin (CTRL AB). D): RASSF1A expression markedly increased sensitivity to apoptosis induction by Apo2L/TRAIL in ACHN cells. Normal kidney epithelial (NKE) cells, even after pretreatment with 5-AZA-dC (200 nM) over 4 days, which alone resulted in moderate apoptosis induction, remained resistant to the apoptosis inducing effects of Apo2L/TRAIL. TUNEL graphs display the means and standard deviations (error bars) of independent experiments.

Article Snippet: Apo2L / TRAIL was obtained from PeproTech (Rocky Hill, NJ), rabbit polyclonal TRAIL neutralizing antibody and control rabbit immunoglobulin from ProSci (Poway, CA).

Techniques: Construct, Transduction, Virus, Clone Assay, Selection, TUNEL Assay, Expressing, Western Blot, Control