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Figure 4. Triptolide overcomes SMO mutant-induced drug resistance. (A–C) The BODIPY-cyclopamine (5 nM) competitive binding assay was separately examined by fluorescent microscope and flow cytometry, Cyclopamine (5 μM) was used as a positive control. (C) Quantification of flow cytometry following BODIPY-cyclopamine competitive binding assay analysis was exhibited. Scale bar: 100 μm. (D, E) Western blot was used to detect the effect of TPL on the expression of Gli1 and ptch1 in NIH3T3 cells after overexpression of SMO473H, and <t>GDC0449</t> (1 μM) was used to control. All results were derived from three independent repeated experiments and represented by mean ± SD, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. vector; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. BODIPY- cyclopamine or SMO473H group; ns, no significance.
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Figure 4. Triptolide overcomes SMO mutant-induced drug resistance. (A–C) The BODIPY-cyclopamine (5 nM) competitive binding assay was separately examined by fluorescent microscope and flow cytometry, Cyclopamine (5 μM) was used as a positive control. (C) Quantification of flow cytometry following BODIPY-cyclopamine competitive binding assay analysis was exhibited. Scale bar: 100 μm. (D, E) Western blot was used to detect the effect of TPL on the expression of Gli1 and ptch1 in NIH3T3 cells after overexpression of SMO473H, and <t>GDC0449</t> (1 μM) was used to control. All results were derived from three independent repeated experiments and represented by mean ± SD, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. vector; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. BODIPY- cyclopamine or SMO473H group; ns, no significance.
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vismodegib gdc0449  (Adooq Bioscience LLC)
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Figure 4. Triptolide overcomes SMO mutant-induced drug resistance. (A–C) The BODIPY-cyclopamine (5 nM) competitive binding assay was separately examined by fluorescent microscope and flow cytometry, Cyclopamine (5 μM) was used as a positive control. (C) Quantification of flow cytometry following BODIPY-cyclopamine competitive binding assay analysis was exhibited. Scale bar: 100 μm. (D, E) Western blot was used to detect the effect of TPL on the expression of Gli1 and ptch1 in NIH3T3 cells after overexpression of SMO473H, and <t>GDC0449</t> (1 μM) was used to control. All results were derived from three independent repeated experiments and represented by mean ± SD, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. vector; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. BODIPY- cyclopamine or SMO473H group; ns, no significance.
Vismodegib Gdc0449, supplied by Adooq Bioscience LLC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 4. Triptolide overcomes SMO mutant-induced drug resistance. (A–C) The BODIPY-cyclopamine (5 nM) competitive binding assay was separately examined by fluorescent microscope and flow cytometry, Cyclopamine (5 μM) was used as a positive control. (C) Quantification of flow cytometry following BODIPY-cyclopamine competitive binding assay analysis was exhibited. Scale bar: 100 μm. (D, E) Western blot was used to detect the effect of TPL on the expression of Gli1 and ptch1 in NIH3T3 cells after overexpression of SMO473H, and GDC0449 (1 μM) was used to control. All results were derived from three independent repeated experiments and represented by mean ± SD, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. vector; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. BODIPY- cyclopamine or SMO473H group; ns, no significance.

Journal: Aging

Article Title: Triptolide inhibits epithelial ovarian tumor growth by blocking the hedgehog/Gli pathway.

doi: 10.18632/aging.205110

Figure Lengend Snippet: Figure 4. Triptolide overcomes SMO mutant-induced drug resistance. (A–C) The BODIPY-cyclopamine (5 nM) competitive binding assay was separately examined by fluorescent microscope and flow cytometry, Cyclopamine (5 μM) was used as a positive control. (C) Quantification of flow cytometry following BODIPY-cyclopamine competitive binding assay analysis was exhibited. Scale bar: 100 μm. (D, E) Western blot was used to detect the effect of TPL on the expression of Gli1 and ptch1 in NIH3T3 cells after overexpression of SMO473H, and GDC0449 (1 μM) was used to control. All results were derived from three independent repeated experiments and represented by mean ± SD, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. vector; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. BODIPY- cyclopamine or SMO473H group; ns, no significance.

Article Snippet: TPL was a purity of 98% purchased from J&K Scientific (USA) (CAS: 38748-32-2), dissolved in dimethyl sulfoxide (DMSO), and stored at -80° C. Gli antagonist GANT61 was purchased from Sigma-Aldrich (USA) (CAS: 500579-04-4), Smoothened antagonist GDC0449 (CAS: 879085-55-9) and Cyclopamine (CAS: 4449-51-8) and Smoothened agonist SAG (CAS: 912545-86-9) were obtained from selleckChem (USA).

Techniques: Mutagenesis, Competitive Binding Assay, Microscopy, Flow Cytometry, Positive Control, Western Blot, Expressing, Over Expression, Control, Derivative Assay, Plasmid Preparation