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: AGO promotes lysosomal degradation of SDC4-CTF. A , Western blot of SDC1/4 CTF and full-length in SW480 cells treated with AGO (indicated doses, 24 h; n = 3). B , Western blot in SW480 cells treated with 20 μM AGO for 0 to 24 h. C and D , immunofluorescence detection and quantification of SDC4 in SW480 cells treated with 20 μM AGO (24 h; n = 6). E , surface SDC4 measured by FACS using FITC-anti-SDC4. F , Western blot of SDC1/4 CTF and full-length in HCT116 cells treated with AGO (indicated doses, 24 h). G , Western blot of SDC4-CTF in multiple colorectal cancer cells treated with 20 μM AGO (24 h). H , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with cycloheximide (50 μg/ml) along with/without 20 μM AGO for indicated times. I , RT-qPCR of indicated genes in SW480 cells treated with 20 μM AGO (24 h; n = 4). J , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with 20 μM AGO, 2.5 μM GM6001, 10 μM TMI-1, or combinations (24 h). K , Western blot in SW480 cells treated with 20 μM AGO, 5 μM BAY11 to 7082, 10 nM PMA, 1 μg/ml LPS, or 20 μg/ml TNFα (24 h). L and M , live-cell imaging and analysis of SDC4-GFP intensity in 293T cells co-expressing DsRed-Rab5A and SDC4-GFP, with 20 μM AGO treatment (n = 4). O , Western blot of lysosome markers and SDC1/4-CTF in SW480 cells treated with 20 μM AGO, 250 nM BAF1, or combination (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗ p < 0.05, ∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns). AGO, andrographolide; SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; FACS, fluorescence-activated cell sorting; BAF1, Bafilomycin A1.

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, Quantitative RT-PCR, Live Cell Imaging, Expressing, Two Tailed Test, Fluorescence, FACS

Journal: Cell reports

Article Title: Endolysosomal damage surveillance enables rapid inflammasome sensing of pathogens

doi: 10.1016/j.celrep.2025.116002

Figure Lengend Snippet: (A) Cell death (% LDH release) and IL-18 release in wild-type (WT) and CASP4 −/− HeLa cells primed with 10 ng/mL IFN-γ overnight and uninfected (medium) or infected with S. Typhimurium ( S . Tm; MOI of 50 unless otherwise indicated) at 6 h post-infection (p.i.). (B–D) Cell death in HeLa cells transfected with control or indicated siRNAs for 48–72 h, primed with IFN-γ, and then infected with S . Tm for 6 h. (E) Cell death in IFN-γ-primed WT and LGALS8 −/− HeLa cells infected with S . Tm at 6 h p.i. (F) Cell death in IFN-γ-primed WT and LGALS3 −/− HeLa cells infected with S . Tm at 6 h p.i. (G) Cell death in HeLa cells transfected with control or indicated siRNAs for 48–72 h, primed with IFN-γ, and then infected with S . Tm for 6 h. (H) Immunoblots for full-length (FL) and N-terminal domain (NT) GSDMD in the lysates and C-terminal domain (CT) GSDMD in the supernatants of IFN-γ-primed WT and LGALS8 −/− HeLa cells infected with S . Tm at 4 h p.i. (I) IL-18 in the supernatants of IFN-γ-primed WT and LGALS8 −/− HeLa cells infected with S . Tm at 6 h p.i. (J and K) Cell death and IL-18 secretion by HeLa cells of indicated genotypes unprimed or primed with 10 ng/mL IFN-γ overnight and infected with S. Tm (MOIs of 50 and 100) at 6 h p.i. (L) Cell death in IFN-γ-primed WT and LGALS8 −/− HeLa cells infected with WT or ΔsifA mutant S . Tm at 6 h p.i. (M and N) Cell death (M) and IL-18 secretion (N) by IFN-γ-primed WT and LGALS8 −/− HeLa cells infected with S . Tm at indicated time points p.i. (O) Immunoblotting of the lysates of WT HeLa cells, LGALS8 −/− HeLa cells, and LGALS8 −/− HeLa cells stably expressing FLAG-galectin-8 for indicated proteins. (P and Q) Cell death (P) and IL-18 secretion (Q) by IFN-γ-primed WT HeLa cells, LGALS8 −/− HeLa cells, and LGALS8 −/− HeLa cells stably expressing FLAG-galectin-8 infected with S . Tm at 6 h p.i. Combined data from 2 (D, J, K, and N) or 3 (A–C, E–G, I, L, M, P, and Q) experiments or one experiment representative of two (H and O) are shown. Data (biological replicates) are plotted in the graphs as mean ± SEM. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; two-way ANOVA with Sidak’s (A–D, G, L–N, and P) or Tukey’s (E, F, I–K, and Q) multiple comparison test. See also and .

Article Snippet: Mouse monoclonal anti- Salmonella Typhimurium LPS , Bio-Rad , Clone 1E6, Cat# 8210-0407; RRID:AB_619235.

Techniques: Infection, Transfection, Control, Western Blot, Mutagenesis, Stable Transfection, Expressing, Comparison

Journal: bioRxiv

Article Title: The Chlamydia trachomatis secreted effector protein CT181 binds to Mcl-1 to prolong neutrophil survival

doi: 10.1101/2025.03.16.643443

Figure Lengend Snippet: ( A ) A2EN or HeLa cells were infected at a MOI of 2.5 with wild-type, tmeA-lx , CT181:: bla , or CT144:: bla . At 48h, cells were lysed and replated on fresh HeLa cell monolayers and infectious forming units were quantified by immunofluorescence microscopy. ( B ) A2EN or HeLa cells were infected at a MOI of 2.5 for 60 min with wild-type, tmeA-lx , CT181:: bla , or CT144:: bla . The number of internal bacteria was determine using differential immunostaining. ( C) To measure inclusion size, A2EN cells were infected at an MOI of 2.5 for 24 h. Bacteria were stained with anti-LPS (green), the inclusion membrane was stained with an anti-IncE antibody (red), and DNA was stained with DAPI (blue). Inclusion diameter was measured in ImageJ. ( A-C ) Data are representative of three independent experiments. Statistical significance was determined using One-Way ANOVA with Dunnett’s multiple comparison post-test comparing the mutants to wild-type. ****P<0.0001, ***P<0.001, **P<0.01, *P<0.05.

Article Snippet: LPS antibody (Novus Cat#NBP1-28820) for enumeration of inclusions by immunofluorescence microscopy.

Techniques: Infection, Immunofluorescence, Microscopy, Bacteria, Immunostaining, Staining, Membrane, Comparison

Journal: Cell Reports

Article Title: PD-1 high CXCR5 – CD4 + peripheral helper T cells promote CXCR3 + plasmablasts in human acute viral infection

doi: 10.1016/j.celrep.2022.111895

Figure Lengend Snippet:

Article Snippet: LPS from E. Coli, TLR4 ligand , NOVUS Biologicals , NBP2-25295.

Techniques: Purification, Recombinant, Cell Isolation, Enzyme-linked Immunosorbent Assay, DNA Library Preparation, Software