acetylated nf κb  (Thermo Fisher)

Journal: Frontiers in Bioscience-Landmark

Article Title: Dapagliflozin Attenuates Myocardial Inflammation and Apoptosis after Coronary Microembolization in Rats by Regulating the SIRT1/NF-κB Signaling Pathway

doi: 10.31083/fbl27082

Figure Lengend Snippet: Fig. 7. DAPA regulates CME-induced myocardial injury through the SIRT1/ NF-κB signaling pathway. (A) Myocardial protein levels of NF-κB p65, Ace-p65, P-p65, and SIRT1 (n = 5). (B) Representative fluorescence images and comparative SIRT1 fluorescence

Article Snippet: Primary antibodies included SIRT1 (1:1000, ab189494, Abcam, Cambridge, UK), TNF-α (1:1000, ab307164, Abcam, Cambridge, UK), Bax (1:1000, ab53154, Abcam, Cambridge, UK), IL-1β (1:1000, ab283818, Abcam, Cambridge, UK), Bcl-2 (1:1000, ab194583, Abcam, Cambridge, UK), NF-κB P65 (1:1000, #8242, CST, Danvers, MA, USA), cleaved caspase-3 (1:1000, #9661, CST, Danvers, MA, USA), Phospho-NF-κB p65 (Ser536) (1:1000, #3033, CST, Danvers, MA, USA), and Acetyl-NF-κB p65 (Lys310) (1:1000, #3045, CST, Danvers, MA, USA). βactin (1:10,000, ab8227, Abcam, Cambridge, UK) served as a reference internal control.

Techniques: Fluorescence

Journal: Foods

Article Title: Role and Mechanism of Sialic Acid in Alleviating Acute Lung Injury through In Vivo and In Vitro Models

doi: 10.3390/foods13182984

Figure Lengend Snippet: SA inhibited the nuclear translocations of NF-κB and AP-1 in LPS-induced HUVEC cells. The protein expressions of transcriptional regulators p65 and c-Jun in nuclear ( A , B ) and cytoplasmic ( C , D ) extracts were examined by western blot analysis. CON: control; LPS: lipopolysaccharide; SA: sialic acid. The images shown here are representative results from three independent experiments. Data are shown as mean ± SD. CON group vs. LPS group or LPS group vs. 25, 50, and 100 μg/mL SA group. * p < 0.05. ** p < 0.01. # p > 0.05.

Article Snippet: Polyclonal antibody tumor necrosis factor-α (TNF-α, CAT. #6945S), interleukin-6 (IL-6, CAT. #12912S), interleukin-1β (IL-1β, CAT. #12242S), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 (CAT. #12629S), activator protein 1 (AP-1, CAT. #9165S), and mitogen-activated protein kinase (MAPK) family antibody (JNK, CAT. #9252S; p-JNK, CAT. #4668S; p38, CAT. #9212S; p-p38, CAT. #4511S) were provided by Cell Signaling Technology (Danver, MA, USA).

Techniques: Western Blot, Control

Journal: Heliyon

Article Title: Acacetin protects against acute lung injury by upregulating SIRT1/ NF-κB pathway

doi: 10.1016/j.heliyon.2024.e37083

Figure Lengend Snippet: Acacetin improved the function of SIRT1 and inhibited the acetylated activation of NF-κB-p65. The levels of NAD + (A), the ratio of NAD + /NADH (B), the mRNA expression of SIRT1 (C), and the activity of SIRT1 (D) were detected in the lung tissue from LPS-treated mice with or without acacetin administration. The representative immunoblots of SIRT1, acetylated NF-κB-p65, and NF-κB-p65 in the nuclear protein, H3 was used as a loading control (E). The levels of SIRT1 (F), acetylated NF-κB-p65 (G), nuclear NF-κB-p65 (H), the ratio of acetylated NF-κB-p65/nuclear NF-κB-p65 (I) were analyzed ( n = 3 independent experiments). The DNA binding activity of NF-κB (J) were assessed ( n = 5 independent experiments). Data were means ± S. D., n = 10 mice/group. ** P < 0.01 vs. control group, ## P < 0.01 vs. LPS group.

Article Snippet: Anti-acetyl-NF-κB-p65 (Lys310, #AF1017), anti-NF-κB-p65 (#BF8005), anti-SIRT1 (#DF6033) and anti-histone H3 (#BF9211) antibodies were obtained from Affinity Biosciences Inc. (Jiangsu, China).

Techniques: Activation Assay, Expressing, Activity Assay, Western Blot, Control, Binding Assay

Journal: Heliyon

Article Title: Acacetin protects against acute lung injury by upregulating SIRT1/ NF-κB pathway

doi: 10.1016/j.heliyon.2024.e37083

Figure Lengend Snippet: Acacetin up-regulated SIRT1 function and restrained NF-κB-p65 acetylated activation in A549 cells. A549 cells were treated with TNF-α (10 ng/ml) in the presence or absence of acacetin (0, 0.3, 1, 3 μM) for 24 h. The levels of NAD + (A), the ratio of NAD + /NADH (B), the mRNA expression of SIRT1 (C), and the activity of SIRT1 (D) were detected. The representative immunoblots of SIRT1, acetylated NF-κB-p65, and NF-κB-p65 in the nuclear protein, H3 was used as a loading control (E). The levels of SIRT1 (F), acetylated NF-κB-p65 (G), nuclear NF-κB-p65 (H), the ratio of acetylated NF-κB-p65/nuclear NF-κB-p65 (I) were analyzed ( n = 3 independent experiments). The DNA binding activity of NF-κB (J) were assessed. Data were means ± S. D., n = 5 independent experiments. ** P < 0.01 vs. control group, ## P < 0.01 vs. TNF-α treated group.

Article Snippet: Anti-acetyl-NF-κB-p65 (Lys310, #AF1017), anti-NF-κB-p65 (#BF8005), anti-SIRT1 (#DF6033) and anti-histone H3 (#BF9211) antibodies were obtained from Affinity Biosciences Inc. (Jiangsu, China).

Techniques: Activation Assay, Expressing, Activity Assay, Western Blot, Control, Binding Assay

Journal: Heliyon

Article Title: Acacetin protects against acute lung injury by upregulating SIRT1/ NF-κB pathway

doi: 10.1016/j.heliyon.2024.e37083

Figure Lengend Snippet: Silencing SIRT1 abolished the efficacy of acacetin for improving SIRT1 function and restraining NF-κB-p65 acetylated activation induced by TNF-α. A549 cells were transfected with Control siRNA or SIRT1 siRNA to further investigate the relationship of acacetin and SIRT1. (A) A549 cells viabilities were measured by MTT assay. (B–E) The contents of LDH, TNF-α, IL-1β, IL-6, and IL-17 in the supernatant of A549 cells were evaluated. (F) The activity of SIRT1 were detected. (G) The representative immunoblots of SIRT1, acetylated NF-κB-p65, and NF-κB-p65 in the nuclear protein, H3 was used as a loading control. The levels of SIRT1 (H), acetylated NF-κB-p65 (I), nuclear NF-κB-p65 (J), and the ratio of acetylated NF-κB-p65/nuclear NF-κB-p65 (K) were analyzed ( n = 3 independent experiments). (L) The DNA binding activity of NF-κB were assessed. Data were means ± S. D., n = 5 independent experiments. ** P < 0.01 vs. control group, ## P < 0.01 vs. TNF-α treated group, $$ P < 0.01 vs. TNF-α+Acacetin group.

Article Snippet: Anti-acetyl-NF-κB-p65 (Lys310, #AF1017), anti-NF-κB-p65 (#BF8005), anti-SIRT1 (#DF6033) and anti-histone H3 (#BF9211) antibodies were obtained from Affinity Biosciences Inc. (Jiangsu, China).

Techniques: Activation Assay, Transfection, Control, MTT Assay, Activity Assay, Western Blot, Binding Assay