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Effects of compound 3b and 4b in migration, cell cycle progression, and MDA accumulation in PCa cells. (a) Representative images of <t>DU145</t> cells migration upon scratch generation in a confluent monolayer. Time point 0 h (T = 0 h) or 60 h (T = 60 h) points post-incubation in presence of DMSO, compound 3b or compound 4b at 20 µM final concentration, are shown. Scale bar = 100 µm. (b) Quantification of scratch area reduction over time normalised to the scratch area at T = 0 h. Wound healing was monitored at 12, 24, 36, 48 and 60 h post-incubation with the indicated compounds. (c) Quantification of cell cycle distribution in DU145 cells treated with DMSO, compound 3b or compound 4b at 20 µM final concentration for 24 h prior to cytofluorimetric analysis. The percentage of cells in each population is shown. (d) Total intracellular ALDH activity measured in DU145 cell lysates following pre-incubation with 20 µM 3b or 4b for 24 h at 37 °C. Cells were extensively washed prior to lysis to eliminate any non-covalently bound material. (e) Quantification of malondialdehyde (MDA) levels in the same cell lysates. MDA concentration was measured and normalised to the total protein content of each sample, as determined by the Bradford assay. In all plots, error bars represent the standard deviation, and asterisks indicate statistical significance levels: p ≤ 0.05 (*), p ≤ 0.01 (**), and p ≤ 0.001 (***).
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Effects of compound 3b and 4b in migration, cell cycle progression, and MDA accumulation in PCa cells. (a) Representative images of <t>DU145</t> cells migration upon scratch generation in a confluent monolayer. Time point 0 h (T = 0 h) or 60 h (T = 60 h) points post-incubation in presence of DMSO, compound 3b or compound 4b at 20 µM final concentration, are shown. Scale bar = 100 µm. (b) Quantification of scratch area reduction over time normalised to the scratch area at T = 0 h. Wound healing was monitored at 12, 24, 36, 48 and 60 h post-incubation with the indicated compounds. (c) Quantification of cell cycle distribution in DU145 cells treated with DMSO, compound 3b or compound 4b at 20 µM final concentration for 24 h prior to cytofluorimetric analysis. The percentage of cells in each population is shown. (d) Total intracellular ALDH activity measured in DU145 cell lysates following pre-incubation with 20 µM 3b or 4b for 24 h at 37 °C. Cells were extensively washed prior to lysis to eliminate any non-covalently bound material. (e) Quantification of malondialdehyde (MDA) levels in the same cell lysates. MDA concentration was measured and normalised to the total protein content of each sample, as determined by the Bradford assay. In all plots, error bars represent the standard deviation, and asterisks indicate statistical significance levels: p ≤ 0.05 (*), p ≤ 0.01 (**), and p ≤ 0.001 (***).
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Effects of compound 3b and 4b in migration, cell cycle progression, and MDA accumulation in PCa cells. (a) Representative images of <t>DU145</t> cells migration upon scratch generation in a confluent monolayer. Time point 0 h (T = 0 h) or 60 h (T = 60 h) points post-incubation in presence of DMSO, compound 3b or compound 4b at 20 µM final concentration, are shown. Scale bar = 100 µm. (b) Quantification of scratch area reduction over time normalised to the scratch area at T = 0 h. Wound healing was monitored at 12, 24, 36, 48 and 60 h post-incubation with the indicated compounds. (c) Quantification of cell cycle distribution in DU145 cells treated with DMSO, compound 3b or compound 4b at 20 µM final concentration for 24 h prior to cytofluorimetric analysis. The percentage of cells in each population is shown. (d) Total intracellular ALDH activity measured in DU145 cell lysates following pre-incubation with 20 µM 3b or 4b for 24 h at 37 °C. Cells were extensively washed prior to lysis to eliminate any non-covalently bound material. (e) Quantification of malondialdehyde (MDA) levels in the same cell lysates. MDA concentration was measured and normalised to the total protein content of each sample, as determined by the Bradford assay. In all plots, error bars represent the standard deviation, and asterisks indicate statistical significance levels: p ≤ 0.05 (*), p ≤ 0.01 (**), and p ≤ 0.001 (***).
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poisson pca  (Virta Labs)
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Effects of compound 3b and 4b in migration, cell cycle progression, and MDA accumulation in PCa cells. (a) Representative images of <t>DU145</t> cells migration upon scratch generation in a confluent monolayer. Time point 0 h (T = 0 h) or 60 h (T = 60 h) points post-incubation in presence of DMSO, compound 3b or compound 4b at 20 µM final concentration, are shown. Scale bar = 100 µm. (b) Quantification of scratch area reduction over time normalised to the scratch area at T = 0 h. Wound healing was monitored at 12, 24, 36, 48 and 60 h post-incubation with the indicated compounds. (c) Quantification of cell cycle distribution in DU145 cells treated with DMSO, compound 3b or compound 4b at 20 µM final concentration for 24 h prior to cytofluorimetric analysis. The percentage of cells in each population is shown. (d) Total intracellular ALDH activity measured in DU145 cell lysates following pre-incubation with 20 µM 3b or 4b for 24 h at 37 °C. Cells were extensively washed prior to lysis to eliminate any non-covalently bound material. (e) Quantification of malondialdehyde (MDA) levels in the same cell lysates. MDA concentration was measured and normalised to the total protein content of each sample, as determined by the Bradford assay. In all plots, error bars represent the standard deviation, and asterisks indicate statistical significance levels: p ≤ 0.05 (*), p ≤ 0.01 (**), and p ≤ 0.001 (***).
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Effects of compound 3b and 4b in migration, cell cycle progression, and MDA accumulation in PCa cells. (a) Representative images of DU145 cells migration upon scratch generation in a confluent monolayer. Time point 0 h (T = 0 h) or 60 h (T = 60 h) points post-incubation in presence of DMSO, compound 3b or compound 4b at 20 µM final concentration, are shown. Scale bar = 100 µm. (b) Quantification of scratch area reduction over time normalised to the scratch area at T = 0 h. Wound healing was monitored at 12, 24, 36, 48 and 60 h post-incubation with the indicated compounds. (c) Quantification of cell cycle distribution in DU145 cells treated with DMSO, compound 3b or compound 4b at 20 µM final concentration for 24 h prior to cytofluorimetric analysis. The percentage of cells in each population is shown. (d) Total intracellular ALDH activity measured in DU145 cell lysates following pre-incubation with 20 µM 3b or 4b for 24 h at 37 °C. Cells were extensively washed prior to lysis to eliminate any non-covalently bound material. (e) Quantification of malondialdehyde (MDA) levels in the same cell lysates. MDA concentration was measured and normalised to the total protein content of each sample, as determined by the Bradford assay. In all plots, error bars represent the standard deviation, and asterisks indicate statistical significance levels: p ≤ 0.05 (*), p ≤ 0.01 (**), and p ≤ 0.001 (***).

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Targeting ALDH7A1 with covalent inhibitors reveals new chemical space for prostate cancer therapy

doi: 10.1080/14756366.2026.2664708

Figure Lengend Snippet: Effects of compound 3b and 4b in migration, cell cycle progression, and MDA accumulation in PCa cells. (a) Representative images of DU145 cells migration upon scratch generation in a confluent monolayer. Time point 0 h (T = 0 h) or 60 h (T = 60 h) points post-incubation in presence of DMSO, compound 3b or compound 4b at 20 µM final concentration, are shown. Scale bar = 100 µm. (b) Quantification of scratch area reduction over time normalised to the scratch area at T = 0 h. Wound healing was monitored at 12, 24, 36, 48 and 60 h post-incubation with the indicated compounds. (c) Quantification of cell cycle distribution in DU145 cells treated with DMSO, compound 3b or compound 4b at 20 µM final concentration for 24 h prior to cytofluorimetric analysis. The percentage of cells in each population is shown. (d) Total intracellular ALDH activity measured in DU145 cell lysates following pre-incubation with 20 µM 3b or 4b for 24 h at 37 °C. Cells were extensively washed prior to lysis to eliminate any non-covalently bound material. (e) Quantification of malondialdehyde (MDA) levels in the same cell lysates. MDA concentration was measured and normalised to the total protein content of each sample, as determined by the Bradford assay. In all plots, error bars represent the standard deviation, and asterisks indicate statistical significance levels: p ≤ 0.05 (*), p ≤ 0.01 (**), and p ≤ 0.001 (***).

Article Snippet: To validate the in vitro and in silico findings in a cellular context, the DU145 PCa cell line was selected as a suitable model due to its high expression of ALDH7A1, as reported in the Human Protein Atlas ( www.proteinatlas.org ).

Techniques: Migration, Incubation, Concentration Assay, Activity Assay, Lysis, Bradford Assay, Standard Deviation