Journal: Frontiers in Immunology

Article Title: MicroRNA-1 Negatively Regulates Peripheral NK Cell Function via Tumor Necrosis Factor-Like Weak Inducer of Apoptosis (TWEAK) Signaling Pathways During PPRV Infection

doi: 10.3389/fimmu.2019.03066

Figure Lengend Snippet: Regulation of NK cell function by miR-1 is mediated by TWEAK signaling. (A) Goat CD16 + CD14 − NK cells were transfected with control miRNA (MC) or miR-1 mimics, or miR-1 mimics and incubated with recombinant TWEAK at the final concentration of 50 nM. Forty-eight hours later, the cells were infected with PPRV at an MOI of 1, and 24 h later, the protein expression of NKG2D, perforin, IFN-γ, and TNF-α was measured by Western blot. Histograms of the right panel showed densitometric analysis of indicated protein normalized to GAPDH to correct for protein loading. (B) Goat CD16 + CD14 − NK cells were transfected with lentivirus TWEAK, si-TWEAK, or respective negative control. Forty-eight hours later, the cells were infected with PPRV at an MOI of 1, and 24 h later, the protein levels of SOCS-1, MyD88, STAT3, pSTAT3, and NF-κB p65 in NK cells were determined by Western blot and normalized to the levels of GAPDH. Results are expressed as means ± standard error of mean (SEM). P -values were calculated using Student's t -test. An asterisk indicates a comparison with the indicated control. * P < 0.05; ** P < 0.01. n.s., not significant.

Article Snippet: For downstream signaling pathway detection, membranes were probed overnight with rabbit anti-SOCS1 (Sangon Biotech, Shanghai, China), rabbit anti-NF-κB p65 (Bioss, Beijing, China), rabbit anti-MyD88 (Bioss, Beijing, China), rabbit anti-STAT3 (Bioss, Beijing, China), or rabbit anti-pSTAT3 (Bioss, Beijing, China).

Techniques: Cell Function Assay, Transfection, Incubation, Recombinant, Concentration Assay, Infection, Expressing, Western Blot, Negative 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 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: Oxidative Medicine and Cellular Longevity

Article Title: Change in Oxidative Stress and Mitochondrial Dynamics in Response to Elevated Cold-Inducible RNA-Binding Protein in Cardiac Surgery-Associated Acute Kidney Injury

doi: 10.1155/2022/3576892

Figure Lengend Snippet: CPB induced mitochondrial dynamics disorder and increased inflammatory factor secretion. (a–f) Expression of IL-6, IL-1 β , and TNF- α in the serum at 0 and 4 hours after CPB. Compared to the CPB group, C23 administration decreased the expression of IL-6, IL-1 β , and TNF- α by 54.0%, 67.5%, and 45.2% at 4 hours, respectively. (g) Western blot analysis of renal Bax, Bcl-2, and cleaved-caspase-3 expression. (h) CPB upregulated the NADPH oxidase via the TLR-4/MyD88 pathway, which promoted the expressions of mitochondrial fission-related proteins Fis1 and Drp1 and reduced the expression of fusion-related protein Mfn2. ∗ P < 0.05 vs. sham; # P < 0.05 vs. CPB.

Article Snippet: The other antibodies used in this study included TLR-4 (19811-1-AP, Proteintech, 1 : 1000), MyD88 (BA2321, Boster, 1 : 500), and anti- β -actin (4967, Cell Signaling Technology, 1 : 1000).

Techniques: Expressing, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Change in Oxidative Stress and Mitochondrial Dynamics in Response to Elevated Cold-Inducible RNA-Binding Protein in Cardiac Surgery-Associated Acute Kidney Injury

doi: 10.1155/2022/3576892

Figure Lengend Snippet: Putative mechanism of CIRP in cardiac surgery-associated acute kidney injury. During cardiac surgery, CIRP is secreted into the circulation in response to hypothermia and hemodynamics change. Extracellular CIRP promotes the expression of NADPH oxidase in renal tubular epithelial cells via the TLR-4/MyD88 pathway and aggravates intracellular oxidative stress. ROS accumulation induces mitochondrial dynamics disorder, which ultimately increases apoptosis and promotes AKI. CIR: cold-inducible RNA-binding protein; NADPH: nicotinamide adenine dinucleotide phosphate; ROS: reactive oxygen species.

Article Snippet: The other antibodies used in this study included TLR-4 (19811-1-AP, Proteintech, 1 : 1000), MyD88 (BA2321, Boster, 1 : 500), and anti- β -actin (4967, Cell Signaling Technology, 1 : 1000).

Techniques: Expressing, RNA Binding Assay

Journal: Pharmacological Research - Modern Chinese Medicine

Article Title: The pharmaceutical applications of total flavonoids extract from Isatis tinctoria L. leaves

doi: 10.1016/j.prmcm.2022.100122

Figure Lengend Snippet: Fig. 8. Effects of TFE from Folium isatidis on the genes expression of TLR4 (a), TRAF6 (b), TRIF (c), I 𝜅B (d) and p65 (e) in lungs tissues of LPS induced ALI mice ( # compared with the control, ∗ compared with LPS, ∗ P < 0.05, ∗ ∗ / ## P < 0.01).

Article Snippet: Antibodies against TLR4, RAF6, TRIF, NF- κB p65, p-NF- κB p65, I κB α, p-I κB α and β-actin were urchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and econdary antibodies were obtained from Wuhan Boster Biological Techology., Ltd (Wuhan, China).

Techniques: Expressing, Control

Journal: Pharmacological Research - Modern Chinese Medicine

Article Title: The pharmaceutical applications of total flavonoids extract from Isatis tinctoria L. leaves

doi: 10.1016/j.prmcm.2022.100122

Figure Lengend Snippet: Fig. 9. Effects of TFE from Folium isatidis on the proteins expression of TLR4 (a), TRAF6 (b), TRIF (c), I 𝜅B (d), p-I 𝜅B (e), p65 (f) and p-p65 (g) in lungs tissues of LPS induced ALI mice ( # compared with the cntrol, ∗ compared with LPS, ∗ P < 0.05, ∗ ∗ / ## P < 0.01).

Article Snippet: Antibodies against TLR4, RAF6, TRIF, NF- κB p65, p-NF- κB p65, I κB α, p-I κB α and β-actin were urchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and econdary antibodies were obtained from Wuhan Boster Biological Techology., Ltd (Wuhan, China).

Techniques: Expressing

Journal: Pharmacological Research - Modern Chinese Medicine

Article Title: The pharmaceutical applications of total flavonoids extract from Isatis tinctoria L. leaves

doi: 10.1016/j.prmcm.2022.100122

Figure Lengend Snippet: Fig. 10. Regulation of TFE from Folium isatidis on TLR4-TRIF-NF- 𝜅B signaling pathway in LPS induced ALI mice.

Article Snippet: Antibodies against TLR4, RAF6, TRIF, NF- κB p65, p-NF- κB p65, I κB α, p-I κB α and β-actin were urchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and econdary antibodies were obtained from Wuhan Boster Biological Techology., Ltd (Wuhan, China).

Techniques: