wild type hct116 cells  (ATCC)

Journal: Biosensors

Article Title: Hybrid Oxygen-Sensing Bio-Scaffolds for 3D Micro-Tissue Models

doi: 10.3390/bios16020122

Figure Lengend Snippet: Dose- and time-dependent profiles of toxicity of the Matrigel–Nano-IR hybrid scaffolds on dispersed HCT116 cells (6k cells in 6 µL aliquots). ( A , B ) show dependence of cell viability on probe concentrations and incubation time, respectively. Liquid scaffold with cells was incubated for 1 h at 37 °C to solidify, then 100 µL of DMEM medium was applied on top. After further incubation (specified), the cells were analysed on a plate reader for total ATP levels using the CellTiter-Glo kit. Results were normalised to the levels of ATP in cells dispersed in Matrigel scaffold without Nano-IR probe.

Article Snippet: Wild-type HCT116 cells were obtained from the American Tissue Culture Collection, ATCC (Manassas, Virginia, VA, USA).

Techniques: Incubation

Journal: Biosensors

Article Title: Hybrid Oxygen-Sensing Bio-Scaffolds for 3D Micro-Tissue Models

doi: 10.3390/bios16020122

Figure Lengend Snippet: Total ATP levels (chemiluminescence counts) for spheroids of HCT116 cells cultured in the hybrid scaffolds containing 0.625 mg/mL Nano-IR probe. CV = 6.83% and 2.84% for spheroids produced by seeding 2k and 4k cells per Lipidure ® coated well. ATP was measured as in .

Article Snippet: Wild-type HCT116 cells were obtained from the American Tissue Culture Collection, ATCC (Manassas, Virginia, VA, USA).

Techniques: Cell Culture, Produced

Journal: Biosensors

Article Title: Hybrid Oxygen-Sensing Bio-Scaffolds for 3D Micro-Tissue Models

doi: 10.3390/bios16020122

Figure Lengend Snippet: Phosphorescent signals for Matrigel–Nano-IR (1 mg/mL) hybrid scaffolds with cultured HCT116 spheroids (2k and 4k, three samples A , C , D in each batch), measured on a wide-field PLIM microscope . ( A ): Two-dimensional intensity images; ( B ): representative line profile of the intensity signal across the spheroid; ( C ): local lifetime values in the different regions of the sample. Positions of the samples on the plate are shown (A-1, C-1, …). Pair t -test analysis: p < 0.01 for the lifetime signal in areas distant and adjacent to the spheroid cultured in the Matrigel–Nano-IR scaffold.

Article Snippet: Wild-type HCT116 cells were obtained from the American Tissue Culture Collection, ATCC (Manassas, Virginia, VA, USA).

Techniques: Cell Culture, Microscopy

Journal: Biosensors

Article Title: Hybrid Oxygen-Sensing Bio-Scaffolds for 3D Micro-Tissue Models

doi: 10.3390/bios16020122

Figure Lengend Snippet: Phosphorescence lifetime profiles of the HCT116 cell spheroids seeded in 6 mL of 50% Matrigel scaffold containing 1 mg/mL of NanO2 probe and covered with 100 mL of McCoy medium, measured on a Victor2 plate reader in TR-F mode. After temperature equilibration of the plate at 37 °C, 100 μL of oil was applied on top of the sample (at ~20 min) to limit back diffusion of O 2 from air.

Article Snippet: Wild-type HCT116 cells were obtained from the American Tissue Culture Collection, ATCC (Manassas, Virginia, VA, USA).

Techniques: Diffusion-based Assay

Journal: bioRxiv

Article Title: MCL1 may not mediate chemoresistance

doi: 10.1101/2025.08.22.671824

Figure Lengend Snippet: MCL1 KO/KD cell line and CST #5453 anti-MCL1 antibody validation by western blot in A ). HCT116, B ). HeLa, C ). HEK293 and D ). MEFs and by IF microscopy in E ). HEK293 cells. SCR = scramble siRNA (negative control). IF images were taken at 40x magnification. Data are representative of three independent experiments.

Article Snippet: Wild-type (WT) cell lines HCT116, HeLa, HEK293, and mouse embryonic fibroblasts (MEFs) were obtained from ATCC through the Cell Services Core Facility, Cleveland Clinic Foundation.

Techniques: Biomarker Discovery, Western Blot, Microscopy, Negative Control

Journal: bioRxiv

Article Title: MCL1 may not mediate chemoresistance

doi: 10.1101/2025.08.22.671824

Figure Lengend Snippet: Summary of results from chemosensitivity studies on WT vs. MCL1 KO/KD cells in both 96-well and 384-well plate formats. One-way ANOVA with multiple comparisons of pGI50 values for each WT and MCL1 KO/KD pair are shown for A) . HCT116, B) . HeLa and C) . MEFs as a result of 48-h chemotherapy treatment in 96-well plates. Multiplicity adjusted P values were obtained for each comparison to determine statistical significance; P > 0.05 (ns), P ≤ 0.05 (*), P ≤ 0.01 (**); ns = not significant. Three pGI50 values were extracted for each WT and MCL1 KO/KD cell line treated with the five chemotherapies since cell viability assays were performed in biological triplicate. SCR = scramble siRNA (negative control). Heatmaps depicting the comparison of mean AUC values for WT and MCL1 KO D-G) HCT116 and H-K) . HeLa cell lines treated with dose titrations of PACL, VINC, VINO, DOX or ETOPO in 384-well plates. Areas were obtained under each percent confluence vs. time growth curve shown in Figure S2A-B . AUCs were averaged from treatments performed in technical triplicate.

Article Snippet: Wild-type (WT) cell lines HCT116, HeLa, HEK293, and mouse embryonic fibroblasts (MEFs) were obtained from ATCC through the Cell Services Core Facility, Cleveland Clinic Foundation.

Techniques: Comparison, Negative Control

Journal: bioRxiv

Article Title: MCL1 may not mediate chemoresistance

doi: 10.1101/2025.08.22.671824

Figure Lengend Snippet: Restoration of MCL1 WT and overexpression of MCL1 NLS in MCL1 KO cell lines to study their effects on chemoresistance. A) . Validation of MCL1 WT and MCL1 NLS localization in MCL1 KO HCT116 and HeLa cells by subcellular fractionation. CYTO = cytosolic fraction; NUC = nuclear fraction. B-C) . Normalization and quantitation of nuclear FLAG expression of MCL1 WT and MCL1 NLS in MCL1 KO HCT116 and HeLa cells, respectively. One-way ANOVA with multiple comparisons of pGI50 values for MCL1 KO vs. KO cell lines reconstituted with MCL1 WT or MCL1 NLS in D) . HCT116 and E) . HeLa for each chemotherapy treatment. Multiplicity adjusted P values were obtained for each comparison to determine statistical significance; P > 0.05 (ns), P ≤ 0.05 (*), P ≤ 0.01 (**); ns = not significant. Three pGI50 values were extracted for each KO and KO cell lines reconstituted with MCL1 WT or MCL1 NLS treated with the five chemotherapies since cell viability assays were performed in biological triplicate.

Article Snippet: Wild-type (WT) cell lines HCT116, HeLa, HEK293, and mouse embryonic fibroblasts (MEFs) were obtained from ATCC through the Cell Services Core Facility, Cleveland Clinic Foundation.

Techniques: Over Expression, Biomarker Discovery, Fractionation, Quantitation Assay, Expressing, Comparison

Journal: bioRxiv

Article Title: MCL1 may not mediate chemoresistance

doi: 10.1101/2025.08.22.671824

Figure Lengend Snippet: MCL1 nuclear translocation assays in HCT116 and HeLa cells. A) . Subcellular fractionation assessment of MCL1 nuclear translocation in response to overnight (20 hr) doxorubicin (DOX), etoposide (ETOPO), paclitaxel (PACL) and oxaliplatin (OXAL) treatments in HeLa cells. CYTO = cytosolic fraction; NUC = nuclear fraction. B) . Subcellular fractionation assessment of MCL1 nuclear translocation in response to 3 hr etoposide treatments. NUC S = soluble nuclear proteins; NUC CHR = chromatin-bound nuclear proteins. C) . IF evaluation of MCL1 nuclear translocation in response to 20 hr chemotherapy treatments in HCT116 and HeLa cells. IF images were taken at 40x magnification. Data are representative of two independent experiments.

Article Snippet: Wild-type (WT) cell lines HCT116, HeLa, HEK293, and mouse embryonic fibroblasts (MEFs) were obtained from ATCC through the Cell Services Core Facility, Cleveland Clinic Foundation.

Techniques: Translocation Assay, Fractionation

Journal: bioRxiv

Article Title: MCL1 may not mediate chemoresistance

doi: 10.1101/2025.08.22.671824

Figure Lengend Snippet: Proximity biotinylation experimental design and results. A) . Schematic of proximity biotinylation approach with arbitrary MCL1-TurboID fusion construct (figure modified from BioRender ). B). MCL1 TurboID plasmid fusion construct design (75 kDa, 700 aa). C) . Validation of GFP TurboID and MCL1 TurboID nuclear localization in HCT116 and HeLa cells by subcellular fractionation. CYTO = cytosolic fraction; NUC = nuclear fraction. D) . Venn diagram comparing number of newly defined MCL1 interactors between HCT116 and HeLa cells with SAINT SP > 0.8. Box shows the 18 shared MCL1 interactors (both nuclear and cytosolic) between both cell lines. Proximity biotinylation experiments were performed in technical triplicate.

Article Snippet: Wild-type (WT) cell lines HCT116, HeLa, HEK293, and mouse embryonic fibroblasts (MEFs) were obtained from ATCC through the Cell Services Core Facility, Cleveland Clinic Foundation.

Techniques: Construct, Modification, Plasmid Preparation, Biomarker Discovery, Fractionation