Journal: Neuron

Article Title: Zika Virus Protease Cleavage of Host Protein Septin-2 Mediates Mitotic Defects in Neural Progenitors

doi: 10.1016/j.neuron.2019.01.010

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Authentication of cell lines used We used HEK293T cells (sex typed as female) and H9 ESC (sex typed as female) that were obtained directly from ATCC, and thus they were not further authenticated.

Techniques: Virus, Recombinant, Mutagenesis, SYBR Green Assay, shRNA, Software

Journal: Acta Pharmaceutica Sinica. B

Article Title: Transfer learning enhanced graph neural network for aldehyde oxidase metabolism prediction and its experimental application

doi: 10.1016/j.apsb.2023.10.008

Figure Lengend Snippet: Identification of hAOX substrates using a liver cytosol assay (oxidative transformation).

Article Snippet: The human liver cytosol (mixed sex; a pool of 50 donors; catalog no. H0610.C, lot no. 1610027; 30 males and 20 females) was purchased from Sekisui XenoTech.

Techniques: Transformation Assay, Inhibition, Positive Control

Journal: Microbial Biotechnology

Article Title: Cutibacterium acnes –Derived Extracellular Vesicles Promote Epithelial Ovarian Cancer Progression by Activating the KEAP1 – NRF2 Antioxidant Pathway to Suppress Ferroptosis

doi: 10.1111/1751-7915.70373

Figure Lengend Snippet: Identification and characterization of pro‐tumorigenic components in C. acnes . (A) Schematic representation of the overall cell experiment workflow. (B) ID8 cells were treated with PBS (P), C. acnes (B), or C. acnes culture supernatant (S), and cell viability was measured by CCK‐8 assay at 0, 6, 24 and 48 h. Data are expressed as fold change relative to the corresponding 0 h value for each group. (C) Colony formation assays were performed to evaluate the clonogenic capacity of ID8 cells. (D–G) Cellular invasion and migration were determined by Transwell and wound‐healing assays under the indicated conditions. (H) Apoptosis of ID8 cells was quantified by flow cytometry 48 h after treatment. (I) Schematic diagram depicting the ultracentrifugation‐based protocol for isolating C. acnes –derived extracellular vesicles (CEVs). (J) Size distribution profile of CEVs determined by nanoparticle tracking analysis. (K) Representative transmission electron microscopy image showing the typical morphology of CEVs. Enlarged views of the selected regions are presented at original magnifications ×100 and ×400, with scale bars of 200 μm and 50 μm, respectively. (L–M) CCK‐8 and colony formation assays revealed that CEVs treatment significantly enhanced ID8 cell proliferation and clonogenicity. (N–Q) Transwell and wound‐healing assays demonstrated that CEVs treatment promoted ID8 cell invasion and migration. (R) Flow cytometry analysis showed that CEVs treatment markedly reduced apoptosis in ID8 cells. Experimental groups: P (PBS), S ( C. acnes culture supernatant) and E (CEVs). Data are presented as mean ± SD; p < 0.05, p < 0.01.

Article Snippet: The ID8 ovarian cancer cell line (official name: ID8; RRID: CVCL_IU14; species: Mus musculus ; sex: female; tissue of origin: ovarian surface epithelium) was obtained from Shanghai Fuxiang Biotechnology (Cat# XF1030, Shanghai, China) in May 2019.

Techniques: CCK-8 Assay, Migration, Flow Cytometry, Derivative Assay, Transmission Assay, Electron Microscopy

Journal: Microbial Biotechnology

Article Title: Cutibacterium acnes –Derived Extracellular Vesicles Promote Epithelial Ovarian Cancer Progression by Activating the KEAP1 – NRF2 Antioxidant Pathway to Suppress Ferroptosis

doi: 10.1111/1751-7915.70373

Figure Lengend Snippet: Extracellular vesicles derived from Cutibacterium acnes induce transcriptional reprogramming and suppress ferroptosis in ID8 ovarian cancer cells. (A) Volcano plot showing differentially expressed genes between PBS‐treated (P) and CEVs‐treated (E) ID8 cells ( p < 0.05, fold change ≥ 2). (B) Heatmap of representative ferroptosis‐related genes significantly altered by CEVs treatment. (C) Gene Ontology (GO) enrichment analysis identifying significantly enriched biological processes, with ROS and glutathione metabolic processes being prominently represented. (D–E) Gene set enrichment analysis (GSEA) results showing significant enrichment of glutathione metabolism and fluid shear stress/atherosclerosis pathways in the CEVs‐treated group. (F–G) Quantitative RT‐PCR analysis of ferroptosis‐related genes, demonstrating upregulation of SLC7A11, GCLM, GPX4, NRF2 and FSP1 and downregulation of KEAP1 and ACSL4 following CEVs treatment ( n = 3). (H–O) Western blot analysis confirming corresponding changes in protein expression of ferroptosis regulators and antioxidant defence components ( n = 3). (P–U) Evaluation of oxidative stress–related indicators showing markedly reduced levels of MDA and intracellular ROS, accompanied by increased activities of SOD and GSH‐Px, elevated GSH content and a higher GSH/GSSG ratio in CEVs‐treated cells, collectively indicating an enhanced antioxidant capacity ( n = 3). (V) Immunofluorescence staining demonstrating increased GPX4 expression in CEVs‐treated ID8 cells (scale bar = 50 μm). Data are presented as means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The ID8 ovarian cancer cell line (official name: ID8; RRID: CVCL_IU14; species: Mus musculus ; sex: female; tissue of origin: ovarian surface epithelium) was obtained from Shanghai Fuxiang Biotechnology (Cat# XF1030, Shanghai, China) in May 2019.

Techniques: Derivative Assay, Shear, Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Microbial Biotechnology

Article Title: Cutibacterium acnes –Derived Extracellular Vesicles Promote Epithelial Ovarian Cancer Progression by Activating the KEAP1 – NRF2 Antioxidant Pathway to Suppress Ferroptosis

doi: 10.1111/1751-7915.70373

Figure Lengend Snippet: Induction of ferroptosis mitigates CEVs‐induced progression of EOC cells in vitro. (A) Colony formation assays showing that CEVs treatment markedly enhanced the clonogenic capacity of ID8 cells, whereas co‐treatment with RSL3 largely reversed this effect. (B, C) Transwell invasion assays and flow cytometric apoptosis analysis demonstrating that CEVs significantly promoted cell invasion and suppressed apoptosis, both of which were attenuated upon RSL3 treatment. (D–L) qRT‐PCR and Western blot analyses of key ferroptosis‐related genes and proteins. CEVs treatment upregulated SLC7A11, GCLM, GPX4, NRF2 and FSP1, while downregulating ACSL4 and KEAP1; these molecular changes were reversed by RSL3, indicating a shift toward a ferroptotic phenotype ( n = 3). (M–R) Biochemical analyses demonstrating that RSL3 treatment markedly elevated MDA levels and intracellular ROS, while significantly reducing SOD activity, GSH‐Px activity, GSH content, and the GSH/GSSG ratio, collectively indicating aggravated oxidative stress compared with CEVs treatment alone ( n = 3). (S) Immunofluorescence analysis confirming a pronounced reduction in GPX4 expression in the RSL3‐treated group (scale bar = 50 μm). Experimental groups: P (PBS), E (CEVs), R (RSL3), and ER (CEVs + RSL3). Data are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The ID8 ovarian cancer cell line (official name: ID8; RRID: CVCL_IU14; species: Mus musculus ; sex: female; tissue of origin: ovarian surface epithelium) was obtained from Shanghai Fuxiang Biotechnology (Cat# XF1030, Shanghai, China) in May 2019.

Techniques: In Vitro, Quantitative RT-PCR, Western Blot, Activity Assay, Immunofluorescence, Expressing

Journal: Molecular Cancer Therapeutics

Article Title: The Selective Estrogen Receptor Degrader ZN-c5 Has Broad Antitumor Activity in Wild-Type and Mutant ER-Positive Breast Cancer Models

doi: 10.1158/1535-7163.MCT-25-0315

Figure Lengend Snippet: ZN-c5 is efficacious as a single agent and in combination in multiple breast cancer models ( A ). ERα and PR protein levels examined by Western blot in different breast cancer cell lines. B, ERα degradation at 24 hours after 1 µm of indicated treatment in different breast cancer cell lines. C, MCF-7 tumor-bearing mice were dosed daily with either vehicle or ZN-c5 and or weekly with fulvestrant as a single agent or in combination with palbociclib at indicated doses for 28 days. Tumor growth curve was shown. D, MCF-7 tumor-bearing mice were dosed daily with either vehicle or ZN-c5 as a single agent or in combination with abemaciclib or ribociclib at indicated doses for 28 days. E, T47D tumor-bearing mice were dosed daily with either vehicle or ZN-c5 or weekly with fulvestrant as a single agent or in combination with palbociclib at indicated doses for 28 days. F and G, ZR-75-1 or HCC1428 tumor-bearing mice were dosed daily with either vehicle or ZN-c5 as a single agent or in combination with palbociclib at indicated doses for 28 days. H, MCF-7 tumor-bearing mice were dosed daily with either vehicle or ZN-c5 as a single agent or in combination with alpelisib at indicated doses for 28 days. Statistical significance was evaluated by one-way ANOVA, *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: CAMA-1 (HTB-21, RRID: CVCL_1115; sex: female) was purchased in 2020, T47D (HTB-133, RRID: CVCL_0553; sex: female) was purchased in 2017, ZR-75-1 (CRL-1500, RRID: CVCL_0588; sex: female) and HCC1428 (CRL-2327, RRID: CVCL_125; sex: female) were purchased in 2018, and HCC1500 (CRL-2329, RRID: CVCL_1254; sex: female) was purchased in 2020 from ATCC and cultured according to ATCC recommendation in RPMI-1640 complete medium.

Techniques: Western Blot