Journal: Nature Communications

Article Title: Targeting chromosomally unstable tumors with a selective KIF18A inhibitor

doi: 10.1038/s41467-024-55300-z

Figure Lengend Snippet: a Structure of VLS-1272. b Potency of VLS-1272 in a KIF18A (1-374) biochemical ADP-Glo assay to measure ATPase activity with varied concentrations of ATP. Data presented as mean values from n = 2 biological replicates. c Inhibition of KIF18A (1-374) by VLS-1272 as measured by ADP-Glo assay in the absence or presence of 0.1 mg/ml microtubules. Data presented as individual values from n = 2 biological replicates. d Representative still images from microtubule gliding filament assays with purified KIF18A 1-480-GFP linked to the glass surface in absence (control) or presence of VLS-1272. Arrows indicate the ends of microtubules that move directionally (control) or remain stationary (VLS-1272). e Plot of microtubule velocities measured in gliding filament assays in the presence or absence of VLS-1272. n = 249 (control) and 177 (VLS-1272) microtubules from 3 independent experiments. Data were compared using an unpaired, two-tailed t -test. **** p < 1.0 x 10 -15 . Data points represent individual microtubule velocities, bars indicate mean and standard deviation. f Potency of VLS-1272 at various timepoints from a global progress curve analysis (GPCA). Data presented as mean values -/+ SD from n = 11 biological replicates. g Inhibition of HCC15 cell proliferation after 4-day treatment with VLS-1272 (black) versus 1 day of VLS-1272 treatment followed by a washout and replacement in media without inhibitor for an additional 3 days (pink). Data presented as individual values from n = 2 biological replicates. h Comparison of the K I calculated by GPCA vs. the EC50 from a 2.5-day CellTiter-Glo proliferation assay in JIMT-1 cells from n = 40 compounds. Each dot represents one compound. Compounds with poor permeability are denoted by “x”. Source data are provided in the Source Data file.

Article Snippet: For side-by-side comparison of VLS-1272 and AM-1882, reactions were performed with 1.25 nM KIF18A and 50 uM ATP for 30 min. AM-1882 was purchased from MedChemExpress (KIF18A-IN-1; Cat. No.: HY-145034).

Techniques: Glo Assay, Activity Assay, Inhibition, Purification, Control, Two Tailed Test, Standard Deviation, Comparison, Proliferation Assay, Permeability

Journal: Nature Communications

Article Title: Targeting chromosomally unstable tumors with a selective KIF18A inhibitor

doi: 10.1038/s41467-024-55300-z

Figure Lengend Snippet: a Growth inhibition as measured by CellTiter-Glo of indicated cancer and non-transformed epithelial cell lines after treatment with VLS-1272 for 168 h. Data presented as individual values from n = 2 biological replicates. b Proliferation of MDA-MB-231 cells over-expressing KIF2A (CIN High ) or KIF2B (CIN Low ) treated with VLS-1272 and assessed by CellTiter-Glo after 168 h of treatment. Data presented as individual values from n = 2 biological replicates. c Growth inhibition of primary T cells isolated from healthy donors after stimulation with αCD3/CD28 beads, treated with VLS-1272 and assessed by CellTiter-Glo after 72 h. Data presented as individual values from n = 2 donors. Dead cell (propidium iodide-positive cells) enumeration by live cell imaging in the presence of vehicle or indicated VLS-1272 concentration for 150 h in JIMT-1 cells ( d ) or CAL51 cells ( e ). Data presented as mean -/+ SD from n = 3 biological replicates. f Dead cell quantification by acridine orange/propidium iodide staining following 3-day treatment with the indicated concentrations of VLS-1272 in OVCAR-3 cells. Data presented as mean -/+ SD from n = 3 biological replicates. P values are calculated compared to untreated control and labeled on the corresponding bar, using one-way ANOVA with Dunnett’s multiple comparisons test. g Western blot of cleaved Caspase 3 (MW: 17 kDa) versus full length Caspase 3 (MW: 35 kDa) after 24- and 72 h treatment of CAL51 and OVCAR-3 cells with VLS-1272. One representative experiment of 2 biological replicates is shown. Growth inhibition as measured by CellTiter-Glo of HCC15 ( h ) and HCC1806 cells ( i ) after treatment with the indicated concentration of VLS-1272 with or without 0.5 or 2 nM BAY-1217389 (MPS1 inhibitor) for 3 days. Data presented as mean values -/+ SD from n = 4 biological replicates. Source data are provided in the Source Data file.

Article Snippet: For side-by-side comparison of VLS-1272 and AM-1882, reactions were performed with 1.25 nM KIF18A and 50 uM ATP for 30 min. AM-1882 was purchased from MedChemExpress (KIF18A-IN-1; Cat. No.: HY-145034).

Techniques: Inhibition, Transformation Assay, Expressing, Isolation, Live Cell Imaging, Concentration Assay, Staining, Control, Labeling, Western Blot

Journal: Nature Communications

Article Title: Targeting chromosomally unstable tumors with a selective KIF18A inhibitor

doi: 10.1038/s41467-024-55300-z

Figure Lengend Snippet: Volcano plot depicting the correlation (Spearman’s rho value) versus significance ( p value computed with Spearman’s rho) of each gene effect profile in the DepMap CRISPR ( a ) or RNAi ( b ) dataset with the VLS-1272 response profile across up to 192 cell lines. c Heatmap of the Spearman’s rho values for each pairwise comparison of treatment response across up to 128 cell lines. The labels on each axis indicate the treatment, and the shading in each box indicates the computed rho value for the comparison of perturbation response. Area under the curve (AUC) values were computed from the VLS-1272 treatment response for use in the analyses in ( a – c ). d Growth inhibition as measured by CellTiter-Glo of MDA-MB-231 cells expressing KIF2A or KIF2B after treatment with the indicated concentrations of Volastertib for 168 h. Data presented as individual values from n = 2 biological replicates. Growth inhibition as measured by CellTiter-Glo of CAL51 and JIMT-1 cells after treatment with the indicated concentrations of Irinotecan ( e ) or Doxorubicin (f) for 168 h. Data presented as mean values -/+ SD from n = 4 (e) or n = 3 ( f ) biological replicates. Source data are provided in the Source Data file.

Article Snippet: For side-by-side comparison of VLS-1272 and AM-1882, reactions were performed with 1.25 nM KIF18A and 50 uM ATP for 30 min. AM-1882 was purchased from MedChemExpress (KIF18A-IN-1; Cat. No.: HY-145034).

Techniques: CRISPR, Comparison, Inhibition, Expressing

Journal: Nature Communications

Article Title: Targeting chromosomally unstable tumors with a selective KIF18A inhibitor

doi: 10.1038/s41467-024-55300-z

Figure Lengend Snippet: a Representative immunofluorescence images of HCC1806 cells treated with DMSO (top) or 37.5 nM VLS-1272 (bottom). Scale bar = 5 µm. One representative image from the treatment and control group from two biological replicates is shown. b Quantification of KIF18A mis-localization in mitotic HCC1806 cells measured as the ratio of KIF18A colocalizing with the α-tubulin spindle to KIF18A colocalizing with DNA (DAPI). Colocalization masks were generated in Harmony high-content analysis software. Data presented as violin plots with the mean (solid line) and quartiles (dashed lines) from n = 233-803 mitotic cells. Adjusted p value is calculated compared to untreated control and labeled next to each condition, using one-way ANOVA with Bonferroni’s multiple comparisons test. c Representative fluorescent images of DAPI-stained mitotic DNA in HCC1806 cells treated with DMSO or VLS-1272. Scale bar = 5 µm. One representative image from the treatment and control group from two biological replicates is shown. d Quantification of the area of DAPI staining (µm 2 ) in pHH3-positive cells treated with increasing doses of VLS-1272. The area of the DAPI-stained DNA was calculated in Harmony high-content analysis software. Data presented as violin plots with the mean (solid line) and quartiles (dashed lines) from n = 652–1434 mitotic cells. Adjusted p value is calculated compared to untreated control and labeled next to each condition, using one-way ANOVA with Bonferroni’s multiple comparisons test. e Quantification of mitotic HCC1806 cells identified by positive staining for pHH3 compared to the number of total DAPI-stained nuclei. The mitotic cell population was calculated in Harmony high-content analysis software. f Percentage of total micronuclei to primary nuclei in cells treated with VLS-1272, calculated in Harmony high-content analysis software. e , f Data presented as mean values with individual data points shown from n = 2 biological replicates. Source data are provided in the Source Data file.

Article Snippet: For side-by-side comparison of VLS-1272 and AM-1882, reactions were performed with 1.25 nM KIF18A and 50 uM ATP for 30 min. AM-1882 was purchased from MedChemExpress (KIF18A-IN-1; Cat. No.: HY-145034).

Techniques: Immunofluorescence, Control, Generated, High Content Screening, Software, Labeling, Staining

Journal: Nature Communications

Article Title: Targeting chromosomally unstable tumors with a selective KIF18A inhibitor

doi: 10.1038/s41467-024-55300-z

Figure Lengend Snippet: Tumor volume measurements after administering vehicle or VLS-1272-SDD to ( a ) female SCID Beige mice bearing HCC15 tumor xenografts or ( b ) female Balb/c nude mice bearing OVCAR-3 tumor xenografts ( n = 10 mice per treatment group). Data presented as mean values -/+ SEM. p values of VLS-1272-treated groups compared to vehicle at Day 36 (HCC15) or Day 29 (OVCAR-3) are indicated and determined using one-way ANOVA with Dunnett’s multiple comparison test. Representative fluorescence images of KIF18A alone or together with α-tubulin and DAPI in tumors harvested from mice treated with vehicle or ( c ) 30 mg/kg VLS-1272 in HCC15 or ( e ) 60 mg/kg VLS-1272 in OVCAR-3. Scale bar = 10 µm ( c ) and scale bar = 5 µm ( e ). Quantification of KIF18A mislocalization measured as the ratio of KIF18A fluorescence colocalizing with α-tubulin over the amount of KIF18A colocalizing with DAPI in HCC15 ( d ) and OVCAR-3 ( f ) xenograft tumor sections. Data presented as mean values -/+ SEM from n = 5 mice per treatment group, ( d ) *** p = 0.00082 and ( f ) *** p = 0.00015 using two-tailed unpaired t test. Representative fluorescence micrographs and quantification of DAPI staining of tumors harvested from mice treated with vehicle or ( g , h ) 30 mg/kg VLS-1272 in HCC15 or ( i , j ) 60 mg/kg VLS-1272 in OVCAR-3. Scale bar = 5 µm ( g , i ). Data presented as mean values -/+ SEM from n = 5 mice per treatment group, ( h ) *** p = 0.00012 and ( j ) * p = 0.012 from two-tailed unpaired t -test. k Mitotic cell counts by pHH3 staining of tumor sections from excised OVCAR-3 xenografts. Data presented as mean -/+ SD from n = 3 mice per group. p values are indicated above the bars and determined using one-way ANOVA with Dunnett’s multiple comparison test. Source data are provided in the Source Data file.

Article Snippet: For side-by-side comparison of VLS-1272 and AM-1882, reactions were performed with 1.25 nM KIF18A and 50 uM ATP for 30 min. AM-1882 was purchased from MedChemExpress (KIF18A-IN-1; Cat. No.: HY-145034).

Techniques: Comparison, Fluorescence, Two Tailed Test, Staining

Journal: Journal of Microbiology & Biology Education

Article Title: Student performance and perceptions in a hybrid laboratory model: an exploratory study of interactive virtual simulations and in-person integration in a foundational microbiology course

doi: 10.1128/jmbe.00203-24

Figure Lengend Snippet: Focus group reflection themes

Article Snippet: The VLS platform used was Labster ( https://www.labster.com ), a computer-based laboratory simulation platform with interactive 3D animations.

Techniques: Microscopy

Journal: Royal Society Open Science

Article Title: Cost-effective, green HPLC determination of losartan, valsartan and their nitrosodiethylamine impurity: application to pharmaceutical dosage forms

doi: 10.1098/rsos.220250

Figure Lengend Snippet: Chemical structures of ( a ) LOS, ( b ) VLS and ( c ) NDEA.

Article Snippet: VLS and LOS analytical standards were supplied and certified by EIPICo Pharmaceuticals, Egypt.

Techniques:

Journal: Royal Society Open Science

Article Title: Cost-effective, green HPLC determination of losartan, valsartan and their nitrosodiethylamine impurity: application to pharmaceutical dosage forms

doi: 10.1098/rsos.220250

Figure Lengend Snippet: Gradiant programme for the proposed method for determination of LOS, VLS and NDEA.

Article Snippet: VLS and LOS analytical standards were supplied and certified by EIPICo Pharmaceuticals, Egypt.

Techniques:

Journal: Royal Society Open Science

Article Title: Cost-effective, green HPLC determination of losartan, valsartan and their nitrosodiethylamine impurity: application to pharmaceutical dosage forms

doi: 10.1098/rsos.220250

Figure Lengend Snippet: Linearity results for the determination of LOS, VLS and NDEA using the proposed method.

Article Snippet: VLS and LOS analytical standards were supplied and certified by EIPICo Pharmaceuticals, Egypt.

Techniques:

Journal: Royal Society Open Science

Article Title: Cost-effective, green HPLC determination of losartan, valsartan and their nitrosodiethylamine impurity: application to pharmaceutical dosage forms

doi: 10.1098/rsos.220250

Figure Lengend Snippet: A typical chromatogram of 20 µl injection of standard solutions of (1) NDEA, (2) VLS and (3) LOS (spiked concentration 2.4, 20.0 and 10.0 µg ml −1 , respectively) under proposed chromatographic conditions.

Article Snippet: VLS and LOS analytical standards were supplied and certified by EIPICo Pharmaceuticals, Egypt.

Techniques: Injection, Concentration Assay

Journal: Royal Society Open Science

Article Title: Cost-effective, green HPLC determination of losartan, valsartan and their nitrosodiethylamine impurity: application to pharmaceutical dosage forms

doi: 10.1098/rsos.220250

Figure Lengend Snippet: Accuracy and precision results of LOS, VLS and NDEA under the proposed method.

Article Snippet: VLS and LOS analytical standards were supplied and certified by EIPICo Pharmaceuticals, Egypt.

Techniques: Concentration Assay

Journal: Royal Society Open Science

Article Title: Cost-effective, green HPLC determination of losartan, valsartan and their nitrosodiethylamine impurity: application to pharmaceutical dosage forms

doi: 10.1098/rsos.220250

Figure Lengend Snippet: Robustness results of LOS, VLS and NDEA under the proposed method.

Article Snippet: VLS and LOS analytical standards were supplied and certified by EIPICo Pharmaceuticals, Egypt.

Techniques: Concentration Assay

Journal: Royal Society Open Science

Article Title: Cost-effective, green HPLC determination of losartan, valsartan and their nitrosodiethylamine impurity: application to pharmaceutical dosage forms

doi: 10.1098/rsos.220250

Figure Lengend Snippet: Comparison between assay results of LOS and VLS in tablet dosage forms using the proposed methods to reported method.

Article Snippet: VLS and LOS analytical standards were supplied and certified by EIPICo Pharmaceuticals, Egypt.

Techniques: Comparison