Journal: Cell Reports Medicine

Article Title: CANDiT: A machine learning framework for differentiation therapy in colorectal cancer

doi: 10.1016/j.xcrm.2025.102421

Figure Lengend Snippet: Summary and working model of how CDX2-reinstatement selectively targets CRC stem cells (A) CRC stem cells, but not normal stem cells, are characterized by enhanced transcription of Wnt/β-catenin and YAP target genes that are established markers of CSCs. Both pathways are known to collaboratively enhance stemness in the colon. , (B) PF treatment activates β1-specific AMPK, reactivates a tumor suppressor pathway (SPS pathway; 1), which attempts to restore tight- (TJ) and adherens (AJ) junctions in the setting of bioenergetic stress. Bioenergetic stress (AMPK activation) and junction restoration reshapes two junction-informed stemness programs, leading to their catastrophic withdrawal: (i) the Hippo-YAP pathway is activated (2), resulting in YAP phosphorylation followed by proteasomal degradation, and consequently, its nuclear exclusion (3) and withdrawal of the YAP/TEAD transcriptional program; (ii) junctional sequestration of β-catenin and consequently, its nuclear exclusion (4) and termination of the β-catenin·TCF/LEF transcriptional program. CDX2 -reinstatement, and differentiation programs that accompanies the same, is likely via AMPK-dependent epigenetic events , (5). Once translated into protein, CDX2 is known to complete and displace β-catenin from TCF/LEF complexes (6), which further accentuates the impact of the SPS pathway on the β-catenin·TCF/LEF transcriptional program. (C) Schematic illustrates the cell state-specific impact of CDX2-reinstatement therapy and the impact of restoration of the stress polarity signaling (SPS) pathway. Pre- (top) and post- (bottom) treatment states of cancers (left) and healthy (right) epithelium are shown. Although activation of the SPS pathway enhances differentiation and junction formation in both cell states, the apoptotic fate is selectively seen in CDX2 -low CSCs.

Article Snippet: AMPK β1 Polyclonal antibody , Proteintech , Cat#10308-1-AP, RRID: AB_513239.

Techniques: Activation Assay, Phospho-proteomics

Journal: Chinese Herbal Medicines

Article Title: Ganoderic acid a derivative induces apoptosis of cervical cancer cells by inhibiting JNK pathway

doi: 10.1016/j.chmed.2024.07.002

Figure Lengend Snippet: (A) Target prediction analysis results. (B) Prediction of GaAD19-JNK interaction. (C–E) Western blot analysis of HeLa cells with GaAD19 treatment for 24 h on expression of JNK, p-JNK, ERK, p-ERK, p38, p-p38 (C), AMPK α 1, p-AMPK α 1-S485, p-AMPK α 1-S496, AMPK β 1, p-AMPK β 1-S108, Akt, p-Akt (D), MAP2K4, MAP2K7 (E), and other proteins (means ± SD, n = 3). (F) Effect of GaAD19 on mRNA level of JNK pathway membrane protein receptor in HeLa cells was determined by qPCR (means ± SD, n = 3). ( ** P < 0.01, *** P < 0.001 vs control group, ns indicates no significant difference).

Article Snippet: AMPK β 1 , Proteintech , 10308–1-AP , 1:500.

Techniques: Western Blot, Expressing, Membrane, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: IMM-H007 promotes hepatic cholesterol and triglyceride metabolism by activating AMPK α to attenuate hypercholesterolemia

doi: 10.1016/j.apsb.2025.05.015

Figure Lengend Snippet: The increase in LDLR protein levels induced by IMM-H007 depends on AMPK α 1/2. (A) LDLR protein levels in Apoe −/− mouse livers were determined via immunofluorescence staining, and the mean immunofluorescence intensity (MFI) of the images was quantified. Scale bar = 50 μm. n = 5. (B) HepG2 cells were treated with H007-M1 at the indicated concentrations for 24 h. LDLR protein expression was determined by Western blotting. (C) HepG2 cells were pretreated with H007-M1 (100 μmol/L) for 12 h and then incubated with CHX (100 μmol/L) for 0, 3, or 6 h. LDLR protein expression was determined by Western blot (top), and the half-life was calculated (bottom). (D, E) HepG2 cells were transfected with scrambled siRNA or AMP-activated protein kinase ( Ampk ) a 1 siRNA and treated with H007-M1 (100 μmol/L) for 24 h. LDLR and AMPK α 1 protein expression was subsequently determined via Western blotting (D), and the band density (E) was quantified. (F, G) HepG2 cells were transfected with scrambled siRNA or AMPKA 2 siRNA and treated with H007-M1 (100 μmol/L) for 24 h, after which LDLR and AMPK α 2 protein expression was determined by Western blotting (F), and the band density was quantified (G). (H, I) HepG2 cells were transfected with scrambled siRNA or AMPKG siRNA and treated with H007-M1 (100 μmol/L) for 24 h, after which LDLR and AMPK γ protein expression was determined by Western blotting (H), and the band density was quantified (I). (J, K) HepG2 cells were transfected with scrambled siRNA or AMPKB siRNA and treated with H007-M1 (100 μmol/L) for 24 h. LDLR and AMPK β protein expression levels were subsequently determined via Western blotting (J), and the band density (K) was quantified. Statistical data are presented as mean ± SEM. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001 versus the indicated group ( n = 3).

Article Snippet: Rabbit anti-LDLR (Cat# 10785-1-AP), PCSK9 (Cat# 55206-1-AP), ABCG1 (Cat# 13578-1-AP), AMPK α 2 (Cat# 18167-1-AP), AMPK β (Cat# 10308-1-AP), AMPK γ (Cat# 10290-1-AP), FXR (Cat# 25055-1-AP), CREB1 (Cat# 12208-1-AP) polyclonal antibodies, mouse anti-FOXO1 (Cat# 66457-1), and HRP-conjugated GAPDH monoclonal antibodies (Cat# HRP-60004), TRITC–conjugated Goat Anti-Rabbit IgG (Cat# SA00007-2) were purchased from Proteintech Group Inc. (Chicago, IL, USA).

Techniques: Immunofluorescence, Staining, Expressing, Western Blot, Incubation, Transfection

Journal: Acta Pharmaceutica Sinica. B

Article Title: IMM-H007 promotes hepatic cholesterol and triglyceride metabolism by activating AMPK α to attenuate hypercholesterolemia

doi: 10.1016/j.apsb.2025.05.015

Figure Lengend Snippet: Hepatocyte Ampka 1/2 deficiency blocked the effect of IMM-H007 on hypercholesterolemia. (A) Ampka flox/flox and h Ampka KO mice were fed a high-fat (HF)/high-cholesterol (HC)/bile-salt (BS) diet with or without IMM-H007 (200 mg/kg body weight/day) for 5 weeks. (B) Determination of serum TC. (C) Determination of serum non-HDL-C. (D) Determination of serum HDL-C levels. (E) Determination of serum TG levels. Statistical data are presented as mean ± SEM. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001 versus the indicated group ( n = 5). (F) RNA-seq assay with mouse liver total RNA. Enrichment analysis of the KEGG and REACTOME pathways ( n = 3). (G, H) Protein levels of LDLR (G, scale bar = 100 μm) AMPK α 1/2 (H, scale bar = 200 μm) in mouse liver was determined by immunofluorescent staining.

Article Snippet: Rabbit anti-LDLR (Cat# 10785-1-AP), PCSK9 (Cat# 55206-1-AP), ABCG1 (Cat# 13578-1-AP), AMPK α 2 (Cat# 18167-1-AP), AMPK β (Cat# 10308-1-AP), AMPK γ (Cat# 10290-1-AP), FXR (Cat# 25055-1-AP), CREB1 (Cat# 12208-1-AP) polyclonal antibodies, mouse anti-FOXO1 (Cat# 66457-1), and HRP-conjugated GAPDH monoclonal antibodies (Cat# HRP-60004), TRITC–conjugated Goat Anti-Rabbit IgG (Cat# SA00007-2) were purchased from Proteintech Group Inc. (Chicago, IL, USA).

Techniques: RNA Sequencing, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: IMM-H007 promotes hepatic cholesterol and triglyceride metabolism by activating AMPK α to attenuate hypercholesterolemia

doi: 10.1016/j.apsb.2025.05.015

Figure Lengend Snippet: IMM-H007 upregulates LDLR expression via the AMPK α 1/2–SENP1–IDOL signaling pathway. (A, B) HepG2 cells were posttranslationally transfected with scrambled siRNA, proprotein convertase subtilisin/kexin type 9 ( PCSK9 ) siRNA (A), or inducible degrader of LDLR ( IDOL ) siRNA (B), after which LDLR, PCSK9, and IDOL protein expression was determined by Western blotting. (C) mRNA levels of PCSK9 and IDOL in HepG2 cells after H007-M1 treatment were measured via qRT-PCR ( n = 3). (D) HEK-293T cells were transfected with Flag- IDOL , HA-small ubiquitin-like modifier 1 ( SUMO1 ), or His-ubiquitin-conjugating enzyme ( UBC9 ) for 24 h and then treated with H007-M1 (100 μmol/L) for 24 h. The SUMOylation of Flag- IDOL was determined by an IP assay using Flag beads. (E) HepG2 cells were transfected with scrambled siRNA or SUMO-specific protease 1 ( SENP1 ) siRNA, after which the protein expression of IDOL, SENP1, and LDLR was determined by Western blotting. (F) Cas9-Ctrl and Cas9- AMPKA cells were treated with H007-M1 at the indicated concentrations for 24 h. IDOL, SENP1, and AMPKA protein expression was determined by Western blotting. (G) Protein expression in the livers of the mice in A was determined by Western blotting. (H) mRNA levels of SENP1 in HepG2 cells after H007-M1 treatment or transfection with AMPKA 1/2 siRNA were measured via qRT-PCR ( n = 3). (I) Putative cAMP response element ( CRE) region in the h SENP1 promoter and the mutated sites. (J) HepG2 cells were transfected with the SENP1 promoter or SENP1 - CRE -mut promoter and Renilla (as an internal control) overnight, followed by 100 μmol/L H007-M1 treatment or AMPKA 1/2 overexpression for another 24 h. The activity of firefly and Renilla luciferases in the cellular lysate was determined via a dual-luciferase reporter assay ( n = 8). Statistical data are presented as mean ± SEM. ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001; NS, not significant versus the indicated groups.

Article Snippet: Rabbit anti-LDLR (Cat# 10785-1-AP), PCSK9 (Cat# 55206-1-AP), ABCG1 (Cat# 13578-1-AP), AMPK α 2 (Cat# 18167-1-AP), AMPK β (Cat# 10308-1-AP), AMPK γ (Cat# 10290-1-AP), FXR (Cat# 25055-1-AP), CREB1 (Cat# 12208-1-AP) polyclonal antibodies, mouse anti-FOXO1 (Cat# 66457-1), and HRP-conjugated GAPDH monoclonal antibodies (Cat# HRP-60004), TRITC–conjugated Goat Anti-Rabbit IgG (Cat# SA00007-2) were purchased from Proteintech Group Inc. (Chicago, IL, USA).

Techniques: Expressing, Transfection, Western Blot, Quantitative RT-PCR, Ubiquitin Proteomics, Control, Over Expression, Activity Assay, Luciferase, Reporter Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: IMM-H007 promotes hepatic cholesterol and triglyceride metabolism by activating AMPK α to attenuate hypercholesterolemia

doi: 10.1016/j.apsb.2025.05.015

Figure Lengend Snippet: IMM-H007 promotes hepatic cholesterol metabolism by activating the AMPK α 1–FXR pathway. (A) Heatmap of the differentially expressed genes related to bile acid metabolism in the liver identified via RNA-seq. (B) Cholesterol metabolism-related protein expression in the livers of the mice shown in A was determined by Western blotting. (C) 293T cells were transfected with the farnesoid X receptor ( FXR ) promoter and Renilla (as an internal control) overnight, followed by 100 μmol/L H007-M1 treatment for another 24 h. The activity of firefly and Renilla luciferases in the cellular lysate was determined via a dual-luciferase reporter assay ( n = 5). (D, E) HepG2 cells were transfected with scrambled siRNA or AMPKA1 siRNA and treated with H007-M1 (100 μmol/L) for 24 h. Then, protein expression was determined by Western blotting (D), and the band density (E) was quantified. (F, G) HepG2 cells were transfected with scrambled siRNA or AMPKA2 siRNA and treated with H007-M1 (100 μmol/L) for 24 h. Then, protein expression was determined by Western blotting (F), and the band density was quantified (G) ( n = 3). (H) Hepatic cholesterol quantitative analysis of the mice in A. (I) Hepatic total bile acid (TBA) quantitative analysis of the mice in A ( n = 5). Statistical data are presented as mean ± SEM. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; NS, not significant versus the indicated group.

Article Snippet: Rabbit anti-LDLR (Cat# 10785-1-AP), PCSK9 (Cat# 55206-1-AP), ABCG1 (Cat# 13578-1-AP), AMPK α 2 (Cat# 18167-1-AP), AMPK β (Cat# 10308-1-AP), AMPK γ (Cat# 10290-1-AP), FXR (Cat# 25055-1-AP), CREB1 (Cat# 12208-1-AP) polyclonal antibodies, mouse anti-FOXO1 (Cat# 66457-1), and HRP-conjugated GAPDH monoclonal antibodies (Cat# HRP-60004), TRITC–conjugated Goat Anti-Rabbit IgG (Cat# SA00007-2) were purchased from Proteintech Group Inc. (Chicago, IL, USA).

Techniques: RNA Sequencing, Expressing, Western Blot, Transfection, Control, Activity Assay, Luciferase, Reporter Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: IMM-H007 promotes hepatic cholesterol and triglyceride metabolism by activating AMPK α to attenuate hypercholesterolemia

doi: 10.1016/j.apsb.2025.05.015

Figure Lengend Snippet: Hepatocyte AMPKα 1/2 deficiency blocked the effect of IMM-H007 on lipid accumulation in the liver. (A) Liver photos of the mice in A. (B, C) H&E (B) and ORO (C) staining of liver sections from the mice in A. Scale bar = 100 μm. (D) TG quantitative analysis of total liver lipid extracts from the mice in A ( n = 5). (E) Adipose triglyceride lipase ( ATGL ) mRNA expression in the livers of the mice shown in A was determined by qRT-PCR ( n = 5). (F) ATGL, diacylglycerol O -acyltransferase 1 (DGAT1) protein expression in the livers of the mice in A was determined by Western blotting ( n = 3). (G) Cas9-Ctrl and Cas9- AMPKA HepG2 cells were treated with H007-M1 at the indicated concentrations for 12 h. Protein expression was determined by Western blotting ( n = 3). (H) Schematic diagram of the potential mechanism by which IMM-H007 decreases hepatic lipid accumulation. Statistical data are presented as mean ± SEM. ∗ P < 0.05 versus the indicated groups.

Article Snippet: Rabbit anti-LDLR (Cat# 10785-1-AP), PCSK9 (Cat# 55206-1-AP), ABCG1 (Cat# 13578-1-AP), AMPK α 2 (Cat# 18167-1-AP), AMPK β (Cat# 10308-1-AP), AMPK γ (Cat# 10290-1-AP), FXR (Cat# 25055-1-AP), CREB1 (Cat# 12208-1-AP) polyclonal antibodies, mouse anti-FOXO1 (Cat# 66457-1), and HRP-conjugated GAPDH monoclonal antibodies (Cat# HRP-60004), TRITC–conjugated Goat Anti-Rabbit IgG (Cat# SA00007-2) were purchased from Proteintech Group Inc. (Chicago, IL, USA).

Techniques: Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: Cell Biology and Toxicology

Article Title: Downregulation of MYBL1 in endothelial cells contributes to atherosclerosis by repressing PLEKHM1-inducing autophagy

doi: 10.1007/s10565-024-09873-6

Figure Lengend Snippet: PLEKHM1 might act as a target of MYBL1 to regulate MYBL1 ( A ) Western blot detected the expressions of PLEKHM1, AMPK, GABARAP and β-actin when MYBL1 was overexpressed and the negative control ( n = 4). ( B ) The quantitative analysis of PLEKHM1, AMPK and GABARAP protein expression were shown. ( C ) CHIP assay was used to detect the interaction between MYBL1 and the promoters of GAPDH, PLEKHM1, AMPK and GABARAP genes ( n = 3). ( D ) PLEKHM1 corresponds to DNA binding site information map. ( E ) Luciferase reporter system was performed to examine the combination between PLEKHM1 and MYBL1 ( n = 3). ( F ) Immunohistochemistry was performed on APOE +/+ and APOE. −/− to assess PLEKHM1 protein expression in atherosclerotic plaques ( n = 8). Scale bars = 200 μm or 20 μm. ( G ) Immunofluorescence assay was used to detect whether MYBL1 entered the nucleus ( n = 3). Scale bars = 200 μm or 20 μm. ( H ) Western blot was used to detect the apoptosis of cells with PLEKHM1, AMPK and GABARAP overexpression when MYBL1 was inhibited ( n = 4). ( I ) The quantitative analysis of Cleaved-Caspase3, Bax and Bcl2 protein expression were shown. ( J ) Flow cytometry was used to detect the apoptosis rate of HUVECs ( n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The primary antibodies were listed as follow: MYBL1 (Affinity, AF9007, 1:1000), PLEKHM1 (Proteintech, 16202–1-AP, 1:1000), β-actin (Proteintech, 81115–1-RR, 1:5000), AMPK (Proteintech, 10929–2-AP, 1: 1000), GABARAP (Proteintech, 18723–1-AP, 1:500), Cleaved-Caspase3 (CST, 9664, 1:1000), Bcl-2 (Proteintech, 26593–1-AP, 1:1000), Bax (Proteintech, 50599–2-Ig, 1:1000), hVps41 (Proteintech, 13869–1-AP, 1:500), hVps11 (Proteintech, 19140–1-AP, 1:500), Rab7 (Proteintech, 55469–1-AP, 1:500), LC3 (Novus, NB100-2220, 1:1000) and p62 (CST, 88588, 1:1000).

Techniques: Western Blot, Negative Control, Expressing, Binding Assay, Luciferase, Immunohistochemistry, Immunofluorescence, Over Expression, Flow Cytometry

Journal: Cell Biology and Toxicology

Article Title: Downregulation of MYBL1 in endothelial cells contributes to atherosclerosis by repressing PLEKHM1-inducing autophagy

doi: 10.1007/s10565-024-09873-6

Figure Lengend Snippet: MYBL1 upregulated the function of HUVECs through PLEKHM1-induced autophagy ( A ) CO-IP was used to detect the binding of PLEKHM1 to hVps41, hVps11 and Rab7 ( n = 3). ( B ) CO-IP was used to detect the binding of hVps41 to PLEKHM1, hVps11 and Rab7 ( n = 3). ( C ) Western blot was used to detect the protein expression of PLEKHM1, p62, LC-3 and β-actin in HUVECs after treatments ( n = 4). ( D ) The quantitative analysis of PLEKHM1, p62 and LC-3II were shown. ( E ) Track to autophagy performed by mCherry-GFP-LC3B was shown ( n = 3). Scale bars = 10 μm. ( F ) The location and expression of LAMP1 and LC3B was determined by immunofluorescence ( n = 3). Scale bars = 10 μm. ( G ) The migration of HUVECs was determined by Transwell assay after treatments ( n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The primary antibodies were listed as follow: MYBL1 (Affinity, AF9007, 1:1000), PLEKHM1 (Proteintech, 16202–1-AP, 1:1000), β-actin (Proteintech, 81115–1-RR, 1:5000), AMPK (Proteintech, 10929–2-AP, 1: 1000), GABARAP (Proteintech, 18723–1-AP, 1:500), Cleaved-Caspase3 (CST, 9664, 1:1000), Bcl-2 (Proteintech, 26593–1-AP, 1:1000), Bax (Proteintech, 50599–2-Ig, 1:1000), hVps41 (Proteintech, 13869–1-AP, 1:500), hVps11 (Proteintech, 19140–1-AP, 1:500), Rab7 (Proteintech, 55469–1-AP, 1:500), LC3 (Novus, NB100-2220, 1:1000) and p62 (CST, 88588, 1:1000).

Techniques: Co-Immunoprecipitation Assay, Binding Assay, Western Blot, Expressing, Immunofluorescence, Migration, Transwell Assay