Fig. 2
From: Anti-BCL2 therapy eliminates giant congenital melanocytic nevus by senolytic and immune induction
BCL2 inhibitor significantly suppresses sphere and colony formation of human GCMN and eliminates GCMN cells. a CCK8 assay evaluating the efficacy of five BCL2 inhibitors in treating GCMN cells from five different patients. Cell viability was assessed after 72 h of inhibitor treatment with 10 μM dilution (n = 5). Mean ± SD. b CCK8 assay comparing the efficacy of Venetoclax (left graph) and Trametinib (right graph) in treating GCMN cells from nine different patients. Cell viability was assessed after 72 h of inhibitor treatment, with concentration gradients used to evaluate average IC50 (n = 9). Mean ± SD. c Morphological changes in GCMN cells following 72 h of treatment with DMSO, Trametinib (10 μM), and Venetoclax (10 μM). Scale bar=100μm. d Flow cytometry analysis detecting ROS expression in GCMN cells after 24 h of treatment with DMSO, Trametinib (10 μM), and Venetoclax (10 μM). e Flow cytometry analysis of apoptotic activity in GCMN cells after 72 h of treatment with DMSO, Trametinib (10 μM), and Venetoclax (10 μM). f Percentage of apoptotic cells in GCMN following 72 h of treatment with DMSO, Trametinib, and Venetoclax (n = 5). Mean ± SD. g Cell sphere assay conducted on GCMN cells after 14 days of low-dose (1 μM). Trametinib and Venetoclax treatment. Scale bar=100μm. h Colony formation assay of GCMN cells after 14 days of treatment with low-dose (1 μM) Trametinib and Venetoclax. Scale bar=100μm. i Average diameters of the formed spheres following 7 and 14 days of the cell sphere assay with DMSO, Trametinib, and Venetoclax (n = 3). Mean ± SD. j Average number of colonies formed after 7 and 14 days of colony formation assay with DMSO, Trametinib, and Venetoclax treatment (n = 3). Mean ± SD. k Significantly up-regulated or down-regulated pathways in Venetoclax-treated (10 μM, 72 h) GCMN cells identified by RNA-seq, comparing DMSO treatment. The apoptosis pathway is labeled with red frame. l Significantly up-regulated gene ontology terms in Venetoclax-treated (10 μM, 72 h) GCMN cells identified by RNA-seq, comparing DMSO treatment. m Hematoxylin and eosin (HE) staining demonstrating the efficacy of intralesional injection of Venetoclax (50 mg/kg daily for 7 days) and Trametinib (10 mg/kg daily for 7 days) in the constructed GCMN patient-derived xenograft (PDX) model. Scale bar=100μm. n Count of the cell density and area of melanin in the dermis based on the HE staining after DMSO, Venetoclax, and Trametinib treatment in GCMN PDX model (n = 3). Mean ± SD
