Journal: Frontiers in Cell and Developmental Biology

Article Title: Investigation of the Potential Mechanisms Underlying Nuclear F-Actin Organization in Ovarian Cancer Cells by High-Throughput Screening in Combination With Deep Learning

doi: 10.3389/fcell.2022.869531

Figure Lengend Snippet: High-throughput screening using a small molecule inhibitor library. (A) , A schematic workflow showing the high-throughput screening performed in the OVCA432-nAc-citrine cell line, using TargetMol small molecule inhibitor library and high-content screening followed by a deep learning approach. (B) , Representative images of OVCA432-nAc-citrine cells treated with 2.5 µM BMS3 to inhibit LIM kinases, or UM164 to inhibit Src and MAPK kinases. Cells were fixed and stained after 6 h inhibitor treatment. The images were captured using high-content screening. Green, nAc-citrine. Blue, DAPI. Scale bar, 20 µm. (C) , The deep learning pipeline is composed of two steps: annotation of nuclear F-actin structures and detection of cells with the presence of nuclear F-actin. The output is shown as cells in which nuclear F-actin structures were detected.

Article Snippet: The TargetMol small inhibitor library includes a molecule named BMS3, which has been reported to target LIMK kinase ( ).

Techniques: High Throughput Screening Assay, High Content Screening, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Investigation of the Potential Mechanisms Underlying Nuclear F-Actin Organization in Ovarian Cancer Cells by High-Throughput Screening in Combination With Deep Learning

doi: 10.3389/fcell.2022.869531

Figure Lengend Snippet: Identification of kinases that are involved in nuclear F-actin assembly in ovarian cancer cells. (A) , Summary of the primary screen using 1247 compounds (small molecule inhibitors). The normalized values for OVCA432-nAc-citrine cells treated with individual inhibitors are shown. (B) , KEGG analysis of proteins potentially targeted by the compounds, that reduce the percentage of cells with nuclear F-actin by more than 20%, or increase the percentage of cells with nuclear F-actin by more than 15%. (C) , The effect of 36 inhibitors targeting PI3K-Akt pathway on nuclear F-actin proportion in OVCA432-nAc-citrine cells. BMS3 was used as a positive control. The orange circles and blue squares represent two biological replicates. (D) , Representative images of OVCA432-nAc-citrine cells treated with DMSO, BMS3 or HG-9-91-01. Scale bar, 20 µm.

Article Snippet: The TargetMol small inhibitor library includes a molecule named BMS3, which has been reported to target LIMK kinase ( ).

Techniques: Positive Control

Journal: Cell Death Discovery

Article Title: FMNL2 suppresses cell migration and invasion of breast cancer: a reduction of cytoplasmic p27 via RhoA/LIMK/Cofilin pathway

doi: 10.1038/s41420-022-00964-z

Figure Lengend Snippet: After transfection, MDA-MB-231 and BT549 cells were incubated with ZOL (100 or 70 μM) or BMS3 (20 or 10 μM) for 24 h, respectively, and then performed for indicated assays. A Representative fluorescent images of F-actin cytoskeleton were displayed. Scale bar, 50 μm. B The levels of FMNL2, Vimentin and Snail were measured by western blotting. C The levels of RhoA/LIMK/Cofilin-related molecules were examined by western blotting. D Co-IP analysis was used to determine the relationship between FMNL2 and RhoA. E The activity levels of RhoA were measured. * P < 0.05 versus NC siRNA group, # P < 0.05 versus siFMNL2 group.

Article Snippet: ZOL as Rho inhibitor, BMS3 (MedChemExpress, Monmouth, NJ, USA) as LIMK inhibitor, MG132 (Sigma, St Louis, MO, USA) as proteasome inhibitor, and doxycycline (DOX; Yeasen Biotechnology, Shanghai, China) as gene expression inducer were used in cell treatment.

Techniques: Transfection, Incubation, Western Blot, Co-Immunoprecipitation Assay, Activity Assay