epcam cd32  (Proteintech)

Journal: Journal of Advanced Research

Article Title: Senkyunolide A interrupts TRAF6-HDAC3 interaction to epigenetically suppress c-MYC and attenuate cholestatic liver injury

doi: 10.1016/j.jare.2025.04.002

Figure Lengend Snippet: The anti-fibrotic mechanisms of SenA are confirmed by HDAC3 inhibition and cholangiocyte-targeted delivery of SenA. (a) The experimental design for HDAC3 inhibition. (b) Representative images of H & E staining, Masson trichrome’s staining and EPCAM immunohistochemistry staining were quantified using Image J software. Scale bar = 50 μm. (c) Relative mRNA levels of H19 , Myc , Epcam and Cdk2 in livers treated with BDL, BDL + BG45, BDL + SenA and BDL + BG45 + SenA. Hprt1 was utilized as an internal control. (d) The experimental design for cholangiocyte-targeted delivery of SenA. (e) Representative images of H & E staining, Masson trichrome’s staining and EPCAM immunohistochemistry staining were quantified using Image J software. Scale bar = 50 μm. (f) Relative mRNA levels of Myc , Krt19 , H19 and Pcna in the liver of mice treated with BDL plus Lipo-Ct, Lipo-SenA, and SenA. Hprt1 was utilized as an internal control. (g) Representative immunoblots against COLLAGEN 1, FIBRONECTIN, c-MYC and β-ACTIN in the liver of mice treated with BDL plus Lipo-Ct or Lipo-SenA. *** P < 0.001, vs. Control group; ### P < 0.001, vs. Model group; $ P < 0.05, vs. Model + SenA group (n = 8).

Article Snippet: For immunohistochemical and immunofluorescent staining, sections were incubated with EpCAM antibody (21050–1-AP, proteintech), KRT19 antibody (10712–1-AP, proteintech), Ki67 antibody (27309–1-AP, proteintech) at 4 °C overnight (detail antibodies information were shown in ).

Techniques: Inhibition, Staining, Immunohistochemistry, Software, Control, Western Blot

Journal: Journal of Advanced Research

Article Title: Senkyunolide A interrupts TRAF6-HDAC3 interaction to epigenetically suppress c-MYC and attenuate cholestatic liver injury

doi: 10.1016/j.jare.2025.04.002

Figure Lengend Snippet: SenA alleviates BDL-induced cholestatic liver injury. (a) Flowchart of animal experiments with associated photographic images of each group (n = 8). (b) Serum AST, ALT, TBA and ALP levels. (c) Representative images of H & E staining and Masson trichrome’s staining was quantified using Image J software. Scale bar = 50 μm. (d) Relative mRNA levels of H19 , Col1a1 , Fn1 and Acta2. Hprt1 was utilized as an internal control. (e) Representative images of EPCAM immunohistochemistry staining. Scale bar = 50 μm. (f) co-immunohistochemistry staining of KRT19 with Ki67 were quantified using Image J software. Scale bar = 25 μm. (g) Relative mRNA levels of Pcna , Sphk2 , P21 and P16 in BDL-induced mouse model. Hprt1 was utilized as an internal control. (h) Analysis of genes downregulated by SenA in BDL mice from GEO database. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. Sham or Normal group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs. Model group (n = 8).

Article Snippet: For immunohistochemical and immunofluorescent staining, sections were incubated with EpCAM antibody (21050–1-AP, proteintech), KRT19 antibody (10712–1-AP, proteintech), Ki67 antibody (27309–1-AP, proteintech) at 4 °C overnight (detail antibodies information were shown in ).

Techniques: Staining, Software, Control, Immunohistochemistry

Journal: Journal of Advanced Research

Article Title: Senkyunolide A interrupts TRAF6-HDAC3 interaction to epigenetically suppress c-MYC and attenuate cholestatic liver injury

doi: 10.1016/j.jare.2025.04.002

Figure Lengend Snippet: RNA-sequencing analysis and experimental verification of the inhibitory effects of SenA on hyperproliferated cholangiocytes. (a) Flowchart of mouse primary cholangiocytes isolation with associated photographic images. (b) GO analysis of DEGs between BDL vs . Sham and DEGs between BDL + SenA vs BDL by R software. The size of dots was used to describe the number of genes enriched in corresponding pathways and the color of dots indicates the significance. (c) The relative expression levels of DEGs implicated in cellular proliferation were shown as a heatmap. (d) Relative mRNA levels of Pcna , Epcam , Myc and Sphk2 in mouse primary cholangiocytes. Hprt1 was utilized as an internal control (n = 3). (e) Representative images of CK7 and KI67 co-immunofluorescence staining. (f) The cell cycle was detected by PI staining. Relative mRNA levels of (g) PCNA and SPHK2 and (h) P21 and P16 in HIBECs. Representative immunoblots against (i) PCNA and SPHK2 and (j) P21 and P16 and β-ACTIN in HIBECs were shown. * P < 0.05, *** P < 0.001, vs. Control group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs. Model group (n = 3).

Article Snippet: For immunohistochemical and immunofluorescent staining, sections were incubated with EpCAM antibody (21050–1-AP, proteintech), KRT19 antibody (10712–1-AP, proteintech), Ki67 antibody (27309–1-AP, proteintech) at 4 °C overnight (detail antibodies information were shown in ).

Techniques: RNA Sequencing, Isolation, Software, Expressing, Control, Immunofluorescence, Staining, Western Blot

Journal: Journal of Advanced Research

Article Title: Senkyunolide A interrupts TRAF6-HDAC3 interaction to epigenetically suppress c-MYC and attenuate cholestatic liver injury

doi: 10.1016/j.jare.2025.04.002

Figure Lengend Snippet: SenA inhibits DR by regulating c-MYC-H19-Let-7a signaling in cholangiocytes. (a) Relative mRNA levels of H19 , HPRT1 was utilized as an internal control. (b)The biological process of MYC-associated pathway was shown, LET7A, LIN28A and CDK2 in HIBECs treated with TCA and different dosages of SenA. HPRT1 was utilized as an internal control. (c) Relative mRNA levels of EPCAM, KRT19, PCNA and CDK2 in H19 overexpression-induced HIBECs. (d) Relative mRNA levels of EPCAM, KRT19, CDK2, CDK1 and PCNA in Let-7a inhibitor-induced HIBECs. (e) Relative mRNA levels of KRT19 , PCNA , CDK1 and EPCAM in H19OE and Let-7a inhibitor-induced HIBECs. (f) Relative mRNA levels of MYC in livers and HIBECs. (g) Representative images of c-MYC immunofluorescence staining. (h) Representative immunoblots against c-MYC and β-ACTIN in HIBECs were shown. (i) The positive relationship between the mRNA expression of H19 and MYC. (j) Representative images of MYC immunohistochemistry staining were quantified using Image J software. Scale bar = 50 μm. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. Control group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs. Model group (n = 3).

Article Snippet: For immunohistochemical and immunofluorescent staining, sections were incubated with EpCAM antibody (21050–1-AP, proteintech), KRT19 antibody (10712–1-AP, proteintech), Ki67 antibody (27309–1-AP, proteintech) at 4 °C overnight (detail antibodies information were shown in ).

Techniques: Control, Over Expression, Immunofluorescence, Staining, Western Blot, Expressing, Immunohistochemistry, Software

Journal: Journal of Advanced Research

Article Title: Senkyunolide A interrupts TRAF6-HDAC3 interaction to epigenetically suppress c-MYC and attenuate cholestatic liver injury

doi: 10.1016/j.jare.2025.04.002

Figure Lengend Snippet: SenA enhances HDAC3 activity via inhibiting the interaction between TRAF6 and HDAC3. (a) The relative expression levels of DEGs implicated in histone acetylation were shown as a heatmap. (b) Relative mRNA levels of HDAC1, HDAC2, HDAC3, HDAC8, HDAC4 and HDAC11 in HIBECs treated with TCA. (c) Relative mRNA levels of KAT8, P300, CBP, PCAF and GCNA in HIBECs treated with TCA. (d) HATs activities in HIBECs and livers. (e) HDACs activities in livers. (f) HDACs activities in HIBECs. (g) Relative mRNA levels of MYC, EPCAM and KRT19 in BG45-stimulated HIBECs. HPRT1 was utilized as an internal control. (h) Ubiquitination analysis of HDAC3. (i) The relative expression levels of DEGs implicated in inflammatory response were shown as a heatmap. (j) Relative mRNA levels of Nfkb , Traf3 , Tab1 and Ikbg1 in primary cholangiocytes. Hprt1 was utilized as an internal control. (k) Representative images of TRAF6 (green) and HDAC3 (red) co-immunofluorescent staining were quantified using Image J software. ** P < 0.01, *** P < 0.001, vs. Control group; # P < 0.05, ## P < 0.01, vs. Model group; $ P < 0.05, vs. Model + SenA group (n = 3).

Article Snippet: For immunohistochemical and immunofluorescent staining, sections were incubated with EpCAM antibody (21050–1-AP, proteintech), KRT19 antibody (10712–1-AP, proteintech), Ki67 antibody (27309–1-AP, proteintech) at 4 °C overnight (detail antibodies information were shown in ).

Techniques: Activity Assay, Expressing, Control, Ubiquitin Proteomics, Staining, Software

Journal: Journal of Advanced Research

Article Title: Senkyunolide A interrupts TRAF6-HDAC3 interaction to epigenetically suppress c-MYC and attenuate cholestatic liver injury

doi: 10.1016/j.jare.2025.04.002

Figure Lengend Snippet: The carbonyl group of SenA is critical for its binding to TRAF6. (a) Molecular docking of SenA and TRAF6. (b) SPR analysis of SenA and TRAF6. (c) CETSA analysis of SenA and TRAF6. (d) DARTS analysis of SenA and TRAF6. (e) MST analysis of SenA and TRAF6. (f) Structural modification (S1) of SenA via Grignard reagent reaction. (g) Relative mRNA levels of KRT19, EPCAM, CDK2 and MYC in TCA plus S1, S2 and S3-treated HIBECs. HPRT1 was utilized as an internal control. (h) MST analysis for S1, S2 and S3. *** P < 0.001, vs. Control group; # P < 0.05, ## P < 0.01, ### P < 0.001 vs. Model group (n = 3).

Article Snippet: For immunohistochemical and immunofluorescent staining, sections were incubated with EpCAM antibody (21050–1-AP, proteintech), KRT19 antibody (10712–1-AP, proteintech), Ki67 antibody (27309–1-AP, proteintech) at 4 °C overnight (detail antibodies information were shown in ).

Techniques: Binding Assay, Modification, Control