Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: MG132 inhibits the degradation of SDC4-CTF. A and B , Western blot of HCT116 cells treated with 0.5, 1, 2.5, or 5 μM MG132 for 12 h, or with 2.5 μM MG132 for 0 to 12 h. C , Western blot of full-length SDC1 and SDC4 in HCT116 cells treated with MG132 (0.5–5 μM, 12 h). D and E , immunofluorescence detection and quantification of HCT116 cells transfected with SDC1-GFP or SDC4-GFP, with/without 5 μM MG132 (4 h; n = 6). F , Western blot of multiple colorectal cancer cell lines treated with 10 μM MG132 (12 h). G , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 0.1, 0.5, or 1 μM proteasome inhibitors (Carfilzomib, Ixazomib, Bortezomib) for 24 h. H , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with PD150606 (2, 5, 10 μM) for 12 h. I , quantification of DQ-BSA fluorescence with MG132 or Earle’s balanced salt solution (EBSS) treatment (n = 4). J , quantification of lysosomal activity using LysoTracker with MG132 or EBSS treatment (n = 4). K , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with siRNA knockdown of proteasome subunits PSMD14, PSMD2, USP14, PSMB5, PSMA6. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, syndecan4; CTF, C-terminal transmembrane (TM) fragment; EBSS, Earle’s balanced salt solution.

Article Snippet: Andrographolide (HY-N0191), MG132 (HY-13259), Bortezomib (HY-10227), Carfilzomib (HY-10455), Ixazomib (HY-10453), DAPT (HY-13027), GM6001 (HY-15768), TMI-1 (HY-101448), LY294002 (HY-10108), Rapamycin (HY-10219), Bafilomycin A1 (HY-100558), cycloheximide (HY-12320), BAY 11 to 7082 (HY-13453), PMA (HY-18739), LPS (HY-D1056), and TNFα (HY- P70426 ) were purchased from MedChemExpress.

Techniques: Western Blot, Immunofluorescence, Transfection, Fluorescence, Activity Assay, Knockdown, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: SDC4-CTF is further cleaved by γ-secretase to release the cytoplasmic fragment. A , confocal microscopy of HCT116 cells transfected with SDC4-GFP or SDC4-CTF-GFP. B , schematic of SDC4-GFP deletion mutants, SNP for Signal peptide, ED for ectodomain, TM for TM motif, C1 and C2 for the constant regions, and V for variable region. C , Western blot of HCT116 cells transfected with SDC1-GFP, SDC4-GFP, or SDC4-CTF-GFP, with/without 5 μM MG132 (4 h). D , sequence alignment of Syndecan family TM C-terminal fragments. The alignment was performed using Clustal W, and analysis was done with ESPript 3.0. E , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with presenilin-1 knockdown (siRNA, 72 h). F , Western blot of HCT116 and SW480 cells treated with 10 μM DAPT, 10 μM MG132, or 2.5 μM GM6001 (12 h). G , schematic of SDC4-CTF degradation via lysosome or proteasome pathways. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, Syndecan4; CTF, C-terminal transmembrane fragment.

Article Snippet: Andrographolide (HY-N0191), MG132 (HY-13259), Bortezomib (HY-10227), Carfilzomib (HY-10455), Ixazomib (HY-10453), DAPT (HY-13027), GM6001 (HY-15768), TMI-1 (HY-101448), LY294002 (HY-10108), Rapamycin (HY-10219), Bafilomycin A1 (HY-100558), cycloheximide (HY-12320), BAY 11 to 7082 (HY-13453), PMA (HY-18739), LPS (HY-D1056), and TNFα (HY- P70426 ) were purchased from MedChemExpress.

Techniques: Confocal Microscopy, Transfection, Western Blot, Sequencing, Knockdown, Two Tailed Test

Journal: iScience

Article Title: Andrographolide inhibits the upregulation of SLC19A3 to block the adipogenic differentiation of adipose-derived stem cells

doi: 10.1016/j.isci.2025.114379

Figure Lengend Snippet: Andrographolide inhibits the adipogenic differentiation of ADSCs (A and B) screening for natural products that inhibit FABP4 (A) and LPL (B) expression. (C) Andrographolide prevents the formation of adipocytes. Scale bars, 100 μm. (D) Andrographolide downregulated the expression differentiation markers dose-dependently. (E) Andrographolide abolished the upregulation of FABP4 and PPARγ protein expression. (F) Andrographolide did not affect the growth of ADSCs as measured by CCK-8. Data are expressed as mean ± SEM. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Andrographolide , MedChemExpress (MCE) , Cat#HY-N0191.

Techniques: Expressing, CCK-8 Assay

Journal: iScience

Article Title: Andrographolide inhibits the upregulation of SLC19A3 to block the adipogenic differentiation of adipose-derived stem cells

doi: 10.1016/j.isci.2025.114379

Figure Lengend Snippet: Andrographolide prevents HFD-induced weight increase and adipogenic differentiation (A and B) Body weights of mice fed with HFD increased markedly. Intragastric administration of andrographolide reduced the increase significantly. AP does not change body weight of mice received normal diet, and did not affect the amounts of the food intake. (C) Histology of mice fed with HFD with or without given AP. Scale bars, 100 μm. (D) Weights of visceral, subcutaneous, and brown adipose tissues from HFD- and HFD plus andrographolide-fed mice. (E) Expression of differentiation markers CEBPα, CEBPβ, PPARγ, FABP4, and LPL in visceral, subcutaneous, and brown adipose tissues from HFD- and HFD plus andrographolide-fed mice. Data are expressed as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Andrographolide , MedChemExpress (MCE) , Cat#HY-N0191.

Techniques: Expressing

Journal: iScience

Article Title: Andrographolide inhibits the upregulation of SLC19A3 to block the adipogenic differentiation of adipose-derived stem cells

doi: 10.1016/j.isci.2025.114379

Figure Lengend Snippet: Andrographolide (AP) inhibits the upregulation of thiamine (vitamin B1) transporter SLC19A3 in ADSCs and in fat tissues from mice fed with HFD (A) RNA-seq analysis of ADSCs induced to adipogenic differentiation in the presence and absence of AP. (B) Induction of SLC19A3 during the differentiation of ADSCs ( n = 3). (C) AP prevented the upregulation of SLC19A3 mRNA ( n = 3). (D) AP diminished the increase of SLC19A3 and FABP4 proteins. The numbers under each band represent relative density compared to the controls. (E) Expression of SLC19A3 in visceral and subcutaneous fat tissues from mice fed with HFD or HFD plus AP. Data are expressed as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Andrographolide , MedChemExpress (MCE) , Cat#HY-N0191.

Techniques: RNA Sequencing, Expressing

Journal: iScience

Article Title: Andrographolide inhibits the upregulation of SLC19A3 to block the adipogenic differentiation of adipose-derived stem cells

doi: 10.1016/j.isci.2025.114379

Figure Lengend Snippet: SLC19A3 mediates the adipogenic differentiation-inhibiting action of andrographolide (A and B) Knock-down of SLC19A3 blocks the adipogenic differentiation of ADSCs examined by oil red O staining (Scale bars, 100 μm) (A), SLC19A3 and LPL expression (B). (C and D) Thiamine reversed the effect of andrographolide on AdSC differentiation (Scale bars, 100 μm) (C) and markers expression (D). Data are expressed as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Andrographolide , MedChemExpress (MCE) , Cat#HY-N0191.

Techniques: Knockdown, Staining, Expressing

Journal: iScience

Article Title: Andrographolide inhibits the upregulation of SLC19A3 to block the adipogenic differentiation of adipose-derived stem cells

doi: 10.1016/j.isci.2025.114379

Figure Lengend Snippet: Andrographolide acts on GSK3β (A) GSK3β was dephosphorylated at S9 upon inducing differentiation of ADSCs toward adipocytes. AP prevents the dephosphorylation of GSK3β dose-dependently. (B) In cellular thermal shift assay, AP protected GSK3β from denaturing and increases the amount of protein in the supernatants, indicative of direct binding. (C) Similar to AP, small molecular inhibitor of GSK3β laduviglusib also blocks the adipogenic differentiation of ADSCs as revealed by oil red O staining. Scale bars, 100 μm.

Article Snippet: Andrographolide , MedChemExpress (MCE) , Cat#HY-N0191.

Techniques: De-Phosphorylation Assay, Thermal Shift Assay, Binding Assay, Staining