Journal: bioRxiv

Article Title: Functional and sensitivity profiling of the KIT Mutation Landscape in Melanoma

doi: 10.64898/2026.02.18.706482

Figure Lengend Snippet: KIT mutations shows differential responses towards common KIT inhibitors in vitro. (A) Western Blot analysis revealed changes in cell growth and proliferation pathways upon expression of KIT mutants in MeWO melanoma cells. (B) Upper panel: Monitoring relative growth of melanoma cell spheroids expressing KIT mutants using high-content microscopy. Lower panel: Representative image depicting spheroid size after 8 days of growth in 3D environment. (C) Western Blot demonstrating the expression of KIT in Ba/F3 cells. (D) Assessment of relative growth of Ba/F3 cells expressing different KIT mutants following withdrawal of IL-3 from the culture medium, measured using cell-titre glo assay. (E) Heatmap shows the relative IC50 of Ba/F3 cell with KIT mutants against KIT inhibitors. (F) Violin plot illustrating the IC50 values of MeWo cells expressing KIT mutants when treated with various KIT inhibitors. Results were obtained from three independent experiments. The horizontal line represents the mean, and statistical analysis was performed using one-way ANOVA.

Article Snippet: Small molecule inhibitors (Imatinib, Sunitinib, Nilotinib, Nintedanib, and Ripretinib) were purchased from MedChemExpress, dissolved in DMSO to a 10 mM stock, and stored at −80°C.

Techniques: In Vitro, Western Blot, Expressing, Microscopy, Glo Assay

Journal: bioRxiv

Article Title: Functional and sensitivity profiling of the KIT Mutation Landscape in Melanoma

doi: 10.64898/2026.02.18.706482

Figure Lengend Snippet: In vivo anti-tumor efficacy of KIT inhibitors against MeWo cells expressing KIT WT/mutants in a mouse xenograft model. (A-C) Response of MeWO-KIT-WT, MeWo-KIT p.L576P, and MeWo-KIT-p.N822K mouse xenograft models to Imatinib, Sunitinib, Nilotinib, and Nintedanib, respectively (n = 3 independent experiments). (D-F) Response of MeWO-KIT-WT, MeWo-KIT p.L576P, and MeWo-KIT-p.N822K mouse xenograft models to Ripretinib (n = 3 independent experiments). Fraction of tumor growth represents the change in tumor volume normalized to day 0 of treatment.

Article Snippet: Small molecule inhibitors (Imatinib, Sunitinib, Nilotinib, Nintedanib, and Ripretinib) were purchased from MedChemExpress, dissolved in DMSO to a 10 mM stock, and stored at −80°C.

Techniques: In Vivo, Expressing

Journal: bioRxiv

Article Title: Myeloid PINK1 represses mtDNA release and immune signaling that impacts neuronal pathology in patient-derived idiopathic PD models

doi: 10.64898/2026.01.07.694713

Figure Lengend Snippet: A) Graphical representation of methodology and findings adapted from Gulen et al. to identify genes induced by the innate immune pathway involving STING, herein referred to as STING-responsive genes (SRG). Cells taken from either skin of human healthy donors, which are then irradiated to promote aging, or brains from old mice were treated with STING inhibitor (H-151). RNAseq (validated by qPCR) was then performed to interrogate the expression of inflammatory genes induced through the activation of the STING pathway during aging. Using their combined data, a list of SRG comprising of 108 genes was compiled (full list shown in Supp. Table 2). B) Dot plot indicating the average expression of SRG across all conditions from the single-cell RNAseq dataset of human iPSC-derived peripheral myeloid cells wherein a larger view showcases key proinflammatory cytokines/chemokines and type I interferons within the SRG panel. The size of the dot determines the proportion of cells expressing the particular gene/gene set while the color scale dictates the normalized average expression. C) Representative immunofluorescence staining of macrophages from human (hiPSC-MDM) and mouse (BMDM) origins for DNA (red) and mitochondria (green) using anti-DNA antibody and AlexaFluor 647-labelled MitoTracker or anti-TOMM40 antibody, respectively. Scale bars: 10 μm and 2 μm (inset). The red-positive puncta dissociated from the mitochondria (green signal) was defined as cytosolic DNA in WT and PiKO stimulated cells, as well as non-stimulated controls. Open and closed squares represent data point for human and mouse experiments, respectively. Two-way ANOVA followed by Holm-Sidak’s multiple comparison test was applied. Mean ± SEM, n=5 independent experiment with each experiment comprised of 20-25 images analyzed. D, E) Representative immunoblots of human and mouse macrophages stained with anti-STING and anti-phospho NF-κβ (pNF-κβ) antibodies. β-actin was used as loading control. The relative protein expression of STING and pNF-κβ in WT and PiKO stimulated cells, as well as non-stimulated controls, were quantified and normalized to β-actin. Open and closed squares represent data point for human and mouse experiments, respectively. Two-way ANOVA followed by Tukey’s multiple comparison test was applied. Mean ± SEM, n=5 independent experiments.

Article Snippet: Matured macrophages were replated and co-stimulated with 500 ng/mL Ultrapure lipopolysaccharide (LPS) from E.coli O111:B4 (Invivogen, tlrl-3pelps) and 50 ng/mL mouse recombinant interleukin-1beta (IL-1β) (PeproTech, 211-11B) with or without 8 μg/mL small molecule STING antagonist H-151 (InvivoGen, inh-h151) for 6 h. For the exchange medium experiment, bone marrow-derived macrophages (BMDM) were treated with inflammatory stimuli for 24 h then cells were washed, and conditioned medium (CM) was collected 24 h later.

Techniques: Irradiation, Expressing, Activation Assay, Derivative Assay, Immunofluorescence, Staining, Comparison, Western Blot, Control

Journal: bioRxiv

Article Title: Myeloid PINK1 represses mtDNA release and immune signaling that impacts neuronal pathology in patient-derived idiopathic PD models

doi: 10.64898/2026.01.07.694713

Figure Lengend Snippet: A) Graphical representation of mtDNA-dependent STING/NF-κβ signaling pathway resulting in inflammation, underscoring H-151 as a STING inhibitor. B) Schematic diagram to describe cell fractionation using commercially available buffers and differential centrifugation. Shown are representative immunoblots of cytosolic (cyto) and mitochondrial (mito) fractions isolated from stimulated WT and PiKO BMDM indicating purity of subcellular fractions according to the expression of mitochondrial protein TOMM20 and cytosolic protein GAPDH. β-actin was used as loading control. Both fractions from stimulated BMDM of WT and PiKO mice were assessed for relative gene expression of mtDNA (MT-ND1). Relative ratios of cyto-to-mito MT-ND1 (cyto/mito) in stimulated PiKO with and without H-151 were normalized to corresponding WT groups. Two-way ANOVA followed by Tukey’s multiple comparison test was applied. Mean ± SEM, n=3 mice per group. C) Representative immunoblots of WT and PiKO BMDM stained with anti-pNF-κβ and anti-STING antibodies. β-actin was used as loading control. The relative protein expression of pNF-κβ and STING in stimulated cells with or without H-151, as well as non-stimulated controls, were quantified and normalized to β-actin. Two-way ANOVA followed by Tukey’s multiple comparison test was applied. Mean ± SEM, n=4 mice per group. D) Quantification of inflammatory target genes downstream of STING innate immune pathway (IL6 and IFNB1) by qPCR across all conditions. Twoway ANOVA followed by Tukey’s multiple comparison test was applied. Mean ± SEM, n=6-7 mice per group.

Article Snippet: Matured macrophages were replated and co-stimulated with 500 ng/mL Ultrapure lipopolysaccharide (LPS) from E.coli O111:B4 (Invivogen, tlrl-3pelps) and 50 ng/mL mouse recombinant interleukin-1beta (IL-1β) (PeproTech, 211-11B) with or without 8 μg/mL small molecule STING antagonist H-151 (InvivoGen, inh-h151) for 6 h. For the exchange medium experiment, bone marrow-derived macrophages (BMDM) were treated with inflammatory stimuli for 24 h then cells were washed, and conditioned medium (CM) was collected 24 h later.

Techniques: Cell Fractionation, Centrifugation, Western Blot, Isolation, Expressing, Control, Gene Expression, Comparison, Staining