anti phospho acly  (Cell Signaling Technology Inc)

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Herbal-based Xuebijing injection ameliorated vascular endothelial dysfunction via inhibiting ACLY/MYB/RIG-I axis in sepsis-associated lung injury.

doi: 10.1016/j.phymed.2025.156573

Figure Lengend Snippet: Fig. 4. ACLY promoted RIG-I/NF-κB signaling via facilitating the acetylation of MYB in ECs. (A). Transcription factors of RIG-I predicted in transcriptome sequence data. (B). The acetylation levels of MYB in HUVEC treated with or without XBJ and BMS-303,141 respectively before LPS stimulation were detected using Western blot after IP with MYB antibody. (C). Immunofluorescent staining showing the nuclear localization of MYB in HUVEC pretreated with or without XBJ or BMS-303,141. (D). The constructed plasmids pGL 4.0, pGL4.0-DDX58, pcDNA3.1 and pcDNA3.1(+)-MYB were transferred into 293T cells. The relative luciferase activity was analyzed by Dual Luciferase Reporter Gene Assay Kit (n = 4). (E-G). Before LPS stimulation, HUVEC were transfected with siRNA specific to MYB. The protein levels of RIG-I, phosphorylated p65, E-selectin, VCAM-1, TF and PAI-1 were analyzed by Western blot.

Article Snippet: The primary antibodies used were VCAM-1 (ab134047, Abcam, Cambridge, UK), E-selectin (20,894–1-AP, Proteintech, Shanghai, China; sc-137,054, Santa Cruz, CA, USA), TF (sc374,441, Santa Cruz, CA, USA), PAI-1 (66,261–1, Proteintech), RIG-I (20,566–1, Proteintech), MYB (17,800–1-AP, Proteintech), p65 (10,745–1, Proteintech), Phosphorylated p65 (Ser468, 82,335–1, Proteintech, Shanghai, China), Phosphorylated ACLY (Ser455, 4331, CST, Danvers, MA), ACLY (13,390, CST), ACSS2 (3658, CST), FASN (3180, CST) and ACC1 (4190, CST).

Techniques: Sequencing, Western Blot, Staining, Construct, Luciferase, Activity Assay, Reporter Gene Assay, Transfection

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Herbal-based Xuebijing injection ameliorated vascular endothelial dysfunction via inhibiting ACLY/MYB/RIG-I axis in sepsis-associated lung injury.

doi: 10.1016/j.phymed.2025.156573

Figure Lengend Snippet: Fig. 6. Individual effectiveness of XBJ compounds on ACLY/MYB/RIG-I axis. (A). Chemical structure of Quercetin. (B). The molecular docking of Quercetin with ACLY. (C-E). HUVEC were pretreated with Quercetin (50 μM) before LPS stimulation. The protein levels of phosphorylated ACLY, RIG-I, phosphorylated p65, E- selectin, VCAM-1, TF and PAI-1 were detected using Western blot. (F). Chemical structure of Kaempferol. (G). The molecular docking of Kaempferol with ACLY. (H- J). HUVEC were pretreated with Kaempferol (50 μM) before LPS stimulation. Western Blotting was used to detect the levels of phosphorylated ACLY, RIG-I, phosphorylated p65, E-selectin, VCAM-1, TF and PAI-1. (K-M). The stability of ACLY protein in HUVEC treated with or without Quercetin and Kaempferol at different temperatures was analyzed by CESTA and western blot.

Article Snippet: The primary antibodies used were VCAM-1 (ab134047, Abcam, Cambridge, UK), E-selectin (20,894–1-AP, Proteintech, Shanghai, China; sc-137,054, Santa Cruz, CA, USA), TF (sc374,441, Santa Cruz, CA, USA), PAI-1 (66,261–1, Proteintech), RIG-I (20,566–1, Proteintech), MYB (17,800–1-AP, Proteintech), p65 (10,745–1, Proteintech), Phosphorylated p65 (Ser468, 82,335–1, Proteintech, Shanghai, China), Phosphorylated ACLY (Ser455, 4331, CST, Danvers, MA), ACLY (13,390, CST), ACSS2 (3658, CST), FASN (3180, CST) and ACC1 (4190, CST).

Techniques: Western Blot

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Herbal-based Xuebijing injection ameliorated vascular endothelial dysfunction via inhibiting ACLY/MYB/RIG-I axis in sepsis-associated lung injury.

doi: 10.1016/j.phymed.2025.156573

Figure Lengend Snippet: Fig. 7. Individual effectiveness of XBJ compounds on ACLY/MYB/RIG-I axis. (A). Chemical structure of Ferulic Acid. (B). The molecular docking of Ferulic Acid with ACLY. (C-E). HUVEC were pretreated with Ferulic Acid (100μM) before LPS stimulation. Immunoblotting was used to detect the levels of phosphorylated ACLY, RIG-I, phosphorylated p65, E-selectin, VCAM-1, TF and PAI-1. (F). Chemical structure of Paeoniflorin. (G). The molecular docking of Paeoniflorin with ACLY. (H-J). HUVEC were pretreated with Paeoniflorin (200 μM) before LPS stimulation. The protein levels of phosphorylated ACLY, RIG-I, phosphorylated p65, E-selectin, VCAM-1, TF and PAI-1 were detected using Western blot. (K-M). The stability of ACLY protein in HUVEC treated with or without Ferulic Acid and Paeoniflorin at different temperatures was analyzed by CESTA and western blot.

Article Snippet: The primary antibodies used were VCAM-1 (ab134047, Abcam, Cambridge, UK), E-selectin (20,894–1-AP, Proteintech, Shanghai, China; sc-137,054, Santa Cruz, CA, USA), TF (sc374,441, Santa Cruz, CA, USA), PAI-1 (66,261–1, Proteintech), RIG-I (20,566–1, Proteintech), MYB (17,800–1-AP, Proteintech), p65 (10,745–1, Proteintech), Phosphorylated p65 (Ser468, 82,335–1, Proteintech, Shanghai, China), Phosphorylated ACLY (Ser455, 4331, CST, Danvers, MA), ACLY (13,390, CST), ACSS2 (3658, CST), FASN (3180, CST) and ACC1 (4190, CST).

Techniques: Western Blot

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Herbal-based Xuebijing injection ameliorated vascular endothelial dysfunction via inhibiting ACLY/MYB/RIG-I axis in sepsis-associated lung injury.

doi: 10.1016/j.phymed.2025.156573

Figure Lengend Snippet: Fig. 9. XBJ ameliorated vascular EC dysfunction via inhibiting ACLY/MYB/RIG-I axis in sepsis-associated lung injury. The compounds in XBJ inhibited the activity of ACLY, thereby inhibiting the expression of RIG-I via reducing the acetylation of transcription factor MYB, leading to the alleviation of EC activation and lung injury induced by sepsis.

Article Snippet: The primary antibodies used were VCAM-1 (ab134047, Abcam, Cambridge, UK), E-selectin (20,894–1-AP, Proteintech, Shanghai, China; sc-137,054, Santa Cruz, CA, USA), TF (sc374,441, Santa Cruz, CA, USA), PAI-1 (66,261–1, Proteintech), RIG-I (20,566–1, Proteintech), MYB (17,800–1-AP, Proteintech), p65 (10,745–1, Proteintech), Phosphorylated p65 (Ser468, 82,335–1, Proteintech, Shanghai, China), Phosphorylated ACLY (Ser455, 4331, CST, Danvers, MA), ACLY (13,390, CST), ACSS2 (3658, CST), FASN (3180, CST) and ACC1 (4190, CST).

Techniques: Activity Assay, Expressing, Activation Assay