human shh elisa kit  (Cusabio)

Journal: Scientific Reports

Article Title: SHH pathway inhibition and astrocyte co-culture induce distinct responses in glioblastoma and cancer stem cells

doi: 10.1038/s41598-026-38199-y

Figure Lengend Snippet: Protein expression level comparison (a) Immunofluorescence images of SHH, SMO and GLI1 protein (b) their fluorescence intensity comparison and (c) ELISA results as representation of SHH secretion. The intensity values of SHH and GLI1 proteins varied with cyclopamine treatment. For SMO protein, the intensity values changed in all groups except for the GBMCs and astrocyte co-culture. Notably, in the GBM CSCs group, it was demonstrated that the response of the astrocyte co-culture to cyclopamine treatment altered the intensity values of the proteins. Statistical significance was considered at * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: ELISA analysis was performed using a commercially available human SHH ELISA kit (Cusabio CSB-E12005h) to investigate the paracrine release of SHH.

Techniques: Expressing, Comparison, Immunofluorescence, Fluorescence, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

Journal: Science signaling

Article Title: Hedgehog-induced ZFYVE21 promotes chronic vascular inflammation by activating NLRP3 inflammasomes in T cells

doi: 10.1126/scisignal.abo3406

Figure Lengend Snippet: (A) Frequency of HLA-DR+IFN-γ+ Tmem in IRI-treated EC:T cell cocultures following EC transfection with control or SHH siRNA. 2–4 independent experiments were performed using N=3 separate leukopack donors (B) Schematic of artery xenograft experimental design. SCID/bg mice were engrafted with human arteries subjected ex vivo to IRI conditions or adoptively transferred with human PBMCs then implanted with osmotic pumps delivering the pharmacological inhibitors of SMO (vismodegib) or GLI (GANT61). (C) Circulating PTCH1hiPD-1hi Tmem frequency in xenografted mice. (D and E) Immunofluorescence staining for CD4+ and CD19+ cells (D) and quantitation of luminal area and intimal CD4+ T cells coverage area (E) in human artery xenografts harvested from mice. (F) IFN-γ concertation in the sera of the xenografted mice. (G and H) SCID/bg mice bearing human artery xenografts were adoptively transferred with FACS-sorted PTCHmid (P1), PTCHlo (P2), or PTCHhi (P3) Tmem. Representative images show immunofluorescence staining for CD4, CD19, and Ulex and Masson’s trichrome staining (G). CD4+ and CD19+ cell coverage, MFI of Ulex, and intimal Masson’s trichrome staining area were quantified (H). (I) FACS analysis of PTCH1 in circulating Tmem and serum IFN-γ concentration in SCID/bg mice that were treated with FACS-sorted PTCHmid (P1), PTCHlo (P2), or PTCHhi (P3) Tmem. (J) Masson’s trichrome staining and quantification of intimal area of human artery xenografts from SCID/bg mice that were treated with FACS-sorted PTCHmid (P1), PTCHlo (P2), or PTCHhi (P3) Tmem. Data in (C–F) represent N=3 mice receiving human arteries from 3 donors. Data in (G–J) represent N=5 mice receiving P2 and P1 Tmem, N=6 mice receiving P3 Tmem. Scale bars, 200μm. *indicates p<0.05, ** indicates p<0.005. Two-way ANOVA followed by Tukey’s pairwise comparison was used for statistical comparisons.

Article Snippet: Patient sera were analyzed using EIA for human SHH (R&D Systems, #DSHH00), human IL-1β (ThermoFisher), and human IFN-γ (ThermoFisher).

Techniques: Transfection, Control, Ex Vivo, Immunofluorescence, Staining, Quantitation Assay, Concentration Assay, Comparison

Journal: Science signaling

Article Title: Hedgehog-induced ZFYVE21 promotes chronic vascular inflammation by activating NLRP3 inflammasomes in T cells

doi: 10.1126/scisignal.abo3406

Figure Lengend Snippet: (A) Quantification of pAktSer473 in Tmem pretreated with antibody against CD28 or Smoothened agonist (SAG) prior to stimulation with antibody against CD3. (B) Quantification of pAktSer473 in FACS-sorted PTCHlo, PTCHmid, and PTCHhi Tmem. (C) Quantification of cleaved Casp-1 in Tmem pretreated with SAG or AktVIII prior to CD3 stimulation. (D and E) Quantification of pAktSer473 in Tmem overexpressing ZFYVE21-GFP (D) or expressing ZFYVE21 shRNA (E) and stimulated with antibody against CD3. (F) Pearson correlation between SHH and IL-18 in DGF patient sera (N=7). (G) PTCHhiPD-1hi Tmem frequency in control (–) and DGF (+) PBMCs. (H) IFN-γ and IL-4+ PTCHhiPD-1hi Tmem frequency in control and DGF PBMCs. (I) Quantification of ZFYVE21, pAktSer473, and cleaved casp-1 in PTCHlo, PTCHmid, and PTCHhi Tmem. (J to L) HLA-DR+PTCHhiPD-1hi Tmem frequency in non-autologous Tmem from a healthy donor stimulated with an antibody against CD3 in the presence of CD28-specific antibody, SAG, MCC950, or vismodegib as indicated. DGF or AMR sera (J and K) or control sera (L) were used as indicated. (M) HLA-DR+PTCHhiPD-1hi Tmem frequency in non-autologous Tmem from a healthy donor stimulated with an antibody specific for CD3 in control serum in the presence of SAG or antibodies specific for MOPC or IL-18 as indicated. (N) Working model for an Hh-induced ZFYVE21-Akt-Casp-1 signaling axis in Tmem. Experimental points in (A–E) and (G–M) reflect technical replicates and were repeated 2–3 times using N=6 separate leukopack donors. * indicates p<0.05, ** indicates p<0.01. Two-way ANOVA followed by Tukey’s pairwise comparison (A-C, I-L) and Student’s t-test (D, E, G, H) were used for statistical comparisons.

Article Snippet: Patient sera were analyzed using EIA for human SHH (R&D Systems, #DSHH00), human IL-1β (ThermoFisher), and human IFN-γ (ThermoFisher).

Techniques: Expressing, shRNA, Control, Comparison