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Image Search Results
Journal: Pharmacological research
Article Title: Cucurbitacin B inhibits non-small cell lung cancer in vivo and in vitro by triggering TLR4/NLRP3/GSDMD-dependent pyroptosis.
doi: 10.1016/j.phrs.2021.105748
Figure Lengend Snippet: Fig. 2. CuB induced NLRP3/caspase-1/GSDMD-mediated pyroptosis. a The morphology changes of A549 cells after treatment with CuB (100 nM) for 8 h (Scale bar=25 µm). b The features of pyroptosis in A549 cells were detected by TEM (Scale bar=2 µm). c The expression of GSDMD-FL, GSDMD-N, pro-IL-1β, mature IL-1β, and HMGB1 in A549 cells. d Immunofluorescence detection of the expression of HMGB1 in A549 cells (Scale bar=20 µm). e The EGFP-NLRP3 and OFPSpark-NEK7 fluorescence were examined by immunofluorescence assay (Scale bar=7.5 µm). f Co-IP analysis of the interaction between NEK7 and NLRP3 in A549 cells after treatment with CuB (100 nM) for 24 h in the presence of VX765 (60 µm). g The expression of NLRP3, NEK7, ASC, pro-caspase-1, and cleaved caspase-1 in A549 cells. h, i Immunofluorescence detection of the caspase-1 expression (Scale bar=25 µm) and ASC dot formation in A549 cells (Scale bar=7.5 µm). j Annexin V and 7-AAD double staining assay was used to confirm CuB (100 nM)-induced pyroptosis in A549 cells.
Article Snippet:
Techniques: Expressing, Immunofluorescence, Fluorescence, Co-Immunoprecipitation Assay, Double Staining
Journal: Pharmacological research
Article Title: Cucurbitacin B inhibits non-small cell lung cancer in vivo and in vitro by triggering TLR4/NLRP3/GSDMD-dependent pyroptosis.
doi: 10.1016/j.phrs.2021.105748
Figure Lengend Snippet: Fig. 3. ROS promoted accumulation of pyroptosis-related Tom20. a JC-1 kit was used to examine the MMP levels in A549 cells (Scale bar=25 µm). b, c ROS level was analyzed by flow cytometry in A549 cells (n ≥3, ***P<0.001 vs CuB; ###P<0.001 vs control group). d Effect of NAC on morphological feature changes in A549 cells (Scale bar=25 µm). e Western blotting analysis of the expression of GSDMD-FL, GSDMD-N, pro-IL-1β, mature IL-1β, pro-caspase-1 and cleaved caspase-1 in A549 cells. f Annexin V and 7-AAD double staining assay was used to identify the inhibition of NAC CuB-induced pyroptosis by NAC in A549 cells. g Immunofluorescence detection of the expression of mitochondrial protein Tom20 in A549 cells (Scale bar=10 µm). h The cell viability in Tom 20 knockdown A549 cells (n ≥3, ***P<0.001 vs CuB; ###P<0.001 vs si-Ctrl group).
Article Snippet:
Techniques: Flow Cytometry, Control, Western Blot, Expressing, Double Staining, Inhibition, Immunofluorescence, Knockdown
Journal: Pharmacological research
Article Title: Cucurbitacin B inhibits non-small cell lung cancer in vivo and in vitro by triggering TLR4/NLRP3/GSDMD-dependent pyroptosis.
doi: 10.1016/j.phrs.2021.105748
Figure Lengend Snippet: Fig. 4. Cytosolic calcium accumulation was essential for CuB-induced pyroptosis. a, b Effect of BAPTA-AM (10 μM) or VX765 (60 µm) on Ca2+ levels in A549 cells (n ≥3, **P<0.01, ***P<0.001 vs CuB; ###P<0.001 vs control group). c The process of the increase of Ca2+ levels in A549 cells was detected by using a Bio Tek CYTATION5 (Scale bar=25 µm). d Immunofluorescence assay was used to examine the expression of Ca2+ fluorescence (Scale bar=7.5 µm). e Effect of BAPTA-AM on the LDH release in A549 cells (n ≥3, ***P<0.001 vs CuB; ###P<0.001 vs control group). f Annexin V/7-AAD double staining assay was used to identify the inhibition of CuB-induced pyroptosis by BAPTA-AM in A549 cells. g The expression of GSDMD-FL, GSDMD-N, pro-IL-1β, mature IL-1β, pro-caspase-1 and cleaved caspase-1 in A549 cells.
Article Snippet:
Techniques: Control, Immunofluorescence, Expressing, Fluorescence, Double Staining, Inhibition
Journal: Pharmacological research
Article Title: Cucurbitacin B inhibits non-small cell lung cancer in vivo and in vitro by triggering TLR4/NLRP3/GSDMD-dependent pyroptosis.
doi: 10.1016/j.phrs.2021.105748
Figure Lengend Snippet: Fig. 5. TLR4 activation promoted the CuB-induced pyroptosis. a The protein expression of TLR4 in A549 cells. b The molecular docking data of CuB on TLR4 dimer were analyzed by using autodock software. c CETSA-WB experiment to further confirm that CuB targeted TLR4 proteins. d The expression of GSDMD-FL, GSDMD-N, pro-IL-1β, mature IL-1β, pro-caspase-1, and cleaved caspase-1 in A549 cells. e Annexin V/7-AAD double staining assay was used to identify the inhi bition of CuB-induced pyroptosis by TAK242 in A549 cells. f The protein expression of TLR4 in A549 cells. g, h The cell viability and LDH activity in TLR4 knockdown A549 cells (n ≥3, *** P<0.001 vs CuB; ###P<0.001 vs NC group). i, j Mitochondrial ROS and cytosolic Ca2+ levels were detected by flow cytometry in A549 cells (n ≥3, ***P<0.001 vs CuB; ###P<0.001 vs NC group).
Article Snippet:
Techniques: Activation Assay, Expressing, Software, Double Staining, Activity Assay, Knockdown, Flow Cytometry
Journal: Pharmacological research
Article Title: Cucurbitacin B inhibits non-small cell lung cancer in vivo and in vitro by triggering TLR4/NLRP3/GSDMD-dependent pyroptosis.
doi: 10.1016/j.phrs.2021.105748
Figure Lengend Snippet: Fig. 7. CuB induced lung cancer cells pyroptosis in mouse models. a HE staining of heart, liver, spleen, lung, and kidney tissues in indicated groups (HE, original magnification, 200×, Scale bar=50 µm). b, c The mitochondrial ROS and cytosolic Ca2+ levels in tumor cells were detected by flow cytometry (n ≥3, ***P<0.001 vs model group, ###P<0.001 vs CuB 0.75 mg/kg group). d, e The protein expressions of TLR4, NEK7, NLRP3, ASC, Tom20, GSDMD-FL, GSDMD-N, pro-IL-1β, mature IL- 1β, pro-caspase-1, cleaved caspase-1and HMGB1 in tumor tissues were detected by western blotting.
Article Snippet:
Techniques: Staining, Flow Cytometry, Western Blot