Journal: Molecular & Cellular Proteomics : MCP

Article Title: Proteomics, Transcriptomics, and Phosphoproteomics Reveal the Mechanism of Talaroconvolutin-A Suppressing Bladder Cancer via Blocking Cell Cycle and Triggering Ferroptosis

doi: 10.1016/j.mcpro.2023.100672

Figure Lengend Snippet: TalaA induced bladder cancer cell death through not apoptosis but ferroptosis. A , “living/dead cell” staining for TalaA-treated SW780 cells. The calcein-AM–stained cells in green represented living cells, and PI stained in red represented dead cells. B , the intracellular ROS was detected by H2DCFDA. TalaA induced ROS in SW780 cells, and the ROS could be neutralized by NAC. The TalaA concentration was 10 μM, and NAC concentration was 1 mM in the experiment. C , the cell morphological evidence of NAC rescuing TalaA-induced cell death in SW780 cells. D , cell apoptosis was detected by flow cytometry. Bladder cancer SW780 cells were co-incubated with 0 to 12 μM TalaA for 48 h. Annexin V-FITC/PI was used to test the apoptosis. E , the effect of TalaA on HMOX1, TF, and STEAP3 protein levels was shown in Heatmap. PRT represented TalaA treatment group; PRC represented DMSO control group. Red represented upregulation, and blue represented downregulation. F , the TalaA-caused changes of ferroptosis-related proteins were shown in the schematic diagram. The red boxes represented upregulated proteins and the green boxes represented downregulated proteins. G , 0 to 12 μM TalaA-treated SW780 cells were stained by crystal violet. TalaA induced vacuolation of SW780 cells in a concentration-dependent manner. H , morphological evidence of deferiprone (DFO) rescuing TalaA-induced cells death. The SW780 cells pretreated with 100 μM DFO for 1 h were co-incubated with 12 μM TalaA for 24 h. I , left , cell activity of SW780 cells incubated with DFO for 24 h, ∗ p < 0.05. Right , cell activity of SW780 cells incubated with DFO and TalaA for 24 h, ∗ p < 0.05. DFO rescued TalaA-suppressed SW780 cell activity, ∗ p < 0.05. J , ferroptosis inhibitor ferrostatin-1 (FS-1) rescued TalaA-repressed bladder cancer SW780 cell activity, ∗ p < 0.05. K , the mitochondrial membrane potential detection. Red represented Mito-tracker Red CMXRos-stained mitochondria and blue represented Hoechst 33342-stained nuclei. L , transmission electron micrograph of SW780 cells treated with TalaA. The red arrows represented the mitochondria. DMSO, dimethyl sulfoxide; HMOX1, heme oxygenase 1; NAC, N-Acetyl-L-cysteine; ROS, reactive oxygen species; TalaA, talaroconvolutin-A; TF, transferrin; .

Article Snippet: To determine the effect of TalaA on the mitochondria of bladder cancer cells, a Mito Tracker Red cmxros Kit (c1049b; Beyotime) was used.

Techniques: Staining, Concentration Assay, Flow Cytometry, Incubation, Control, Activity Assay, Membrane, Transmission Assay

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Hedyotis diffusa injection induces ferroptosis via the Bax/Bcl2/VDAC2/3 axis in lung adenocarcinoma.

doi: 10.1016/j.phymed.2022.154319

Figure Lengend Snippet: Fig. 3. HDI-induced ferroptosis in lung adenocarcinoma cells is independent of GPX4 and PUFA-PLS pathways. (A) Overview of mechanisms underlying ferroptosis. (B) Protein expression of GPX4 after transfection with an overexpression plasmid. (C, F) The GPX4 overexpression (OE) plasmid or zileuton were added at the time of HDI treatment to A549 and H1975 cells , followed by staining with BODIPY-C11 and the evaluation of lipid ROS levels using flow cytometry (D, G). A549 and H1975 cells were simultaneously treated with the GPX4 OE plasmid or zileuton with HDI and stained with DAPI for 30 min. Fe2+ was evaluated using a Fe2+ Assay Kit, followed by fluorescence microscopy at 100 × . (E, H) Detection of the lipid oxidation product malondialdehyde. *p < 0.05, **p < 0.01. vs the control group.

Article Snippet: Additionally, a Lipid Peroxidation Malondialdehyde (MDA) Assay Kit (#S0131S) Mito-Tracker Red CMX ROS (#C1049B), and 4′,6-diamidino-2-phenylindole (DAPI) (#C1005) were purchased from Beyotime (Shanghai, China).

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, Staining, Flow Cytometry, Fluorescence, Microscopy, Control

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Hedyotis diffusa injection induces ferroptosis via the Bax/Bcl2/VDAC2/3 axis in lung adenocarcinoma.

doi: 10.1016/j.phymed.2022.154319

Figure Lengend Snippet: Fig. 4. HDI-induced ferroptosis in lung adenocarcinoma cells is associated with VDAC2/3. (A) Protein expression of VDAC2/3 after siRNA transfection, S1, S2, and S3 represent three knockdown sequences, both A549 and H1975 cells were S3 knockdown with the highest efficiency, while the S3 sequences were used for the knockdown of A549 and H1975 cells in subsequent experiments. (B) A549 and H1975 cells were transfected with siRNA targeting VDAC2/3 at the time of HDI treatment and stained with BODIPY-C11, followed by analyses of lipid ROS levels using flow cytometry. (C) VDAC2/3 siRNA transfected at the time of HDI treatment was stained with DAPI for 30 min followed by an analysis of iron using a Fe2+ Assay Kit for 30 min and observation using fluorescence microscopy at 100 × . (D) Detection of the lipid oxidation product malondialdehyde. *p < 0.05, **p < 0.01, vs the control group. ##p < 0.01, vs the HDI group.

Article Snippet: Additionally, a Lipid Peroxidation Malondialdehyde (MDA) Assay Kit (#S0131S) Mito-Tracker Red CMX ROS (#C1049B), and 4′,6-diamidino-2-phenylindole (DAPI) (#C1005) were purchased from Beyotime (Shanghai, China).

Techniques: Expressing, Transfection, Knockdown, Staining, Flow Cytometry, Fluorescence, Microscopy, Control