osmi1 #21894  (Cayman Chemical)

Journal: Neoplasia (New York, N.Y.)

Article Title: Twist1-induced suppression of oncogene-induced senescence in non-small cell lung cancer requires the transactivation domain of Twist1

doi: 10.1016/j.neo.2025.101179

Figure Lengend Snippet: The HBP/O-GlcNAc and MYC axes are required for TWIST1-mediated suppression of OIS. At 72-hours post-adenoviral transduction for HRas G12V -EGFP or control-EGFP in HBEC— Control , HBEC- TWIST1 wt and HBEC- TWIST F187G : Senescence-associated beta-galactosidase staining images and quantification of cells treated 48-hours with ( A-B ) OGA inhibitor TMG (50μ M), ( A-C ) GFPT2 inhibitor DON (5μ M), ( A-D ) OGT inhibitor OSMI1 (7.5μ M), and ( E-F ) MYC inhibitor MYCi975 (0.25μ M or 0.6μ M or 1.2μ M). Data are presented as average values ± S.D. ( n = 3; two-way anova test: P > 0.05(ns)/ P < 0.05(*)/0.01(**)/0.001(***)). [Overall, parental HBEC cells were globally following the same profile as the conditions HBEC— Control, ].

Article Snippet: Cells were treated as previously described [ ] with the pharmacological inhibitors for 48 hours prior to analysis using the following compounds: 5μ M DON (6-Diazo-5-oxo-l-norleucine, #D2141, Sigma-Aldrich (RRID: SCR_008988)), 50μ M TMG (Thiamet-G #4390, Tocris Bioscience (RRID: SCR_003689)), 7.5μ M OSMI1 (#21894, Cayman Chemical (RRID: SCR_008945)), or 0.25μM/0.6μM/1.2μ M MYCi975 (synthesized and provided by Dr Abdulkadir laboratory, Northwestern University, Chicago, IL, USA), [ ].

Techniques: Transduction, Control, Staining

Journal: Scientific Reports

Article Title: O-GlcNAc Transferase Inhibition Differentially Affects Breast Cancer Subtypes

doi: 10.1038/s41598-019-42153-6

Figure Lengend Snippet: OGT inhibition is potently cytotoxic in TNBC cell lines. ( A ) Relative decrease in total protein O-GlcNAcylation, after 4 and 24 hours of treatment with 40 µM OSMI1 in six BC cell lines. Percentage above indicates median decrease compared to the respective DMSO-treated controls, at least 2 independent experiments. ( B ) Relative viability in BC cell lines after 72 hours of treatment with OSMI1. MTS assay. At least 3 biological replicates for each cell line. Error bars - SEM. TNBC lines marked in black and grey; triple-positive cell lines in red. ( C ) EC50 and EC20 concentrations of OSMI1, mean from at least three independent experiments for each cell line. ( D ) OGT transient knock-down in four BC cell lines, representative out of at least three experiments. ( E ) BC cells pictured after 72 hours of treatment with 20 µM OSMI1, or 5 days of transient OGT knockdown. ( F ) Apoptotic population in MDA-MB-231 following 3, 4 and 5 days of transient OGT knock-down, TUNEL assay. Error bars - standard deviation, n = at least 2. *p < 0.05; **p < 0.005; ***p < 0.001 unpaired t-test, comparing knock-down samples to the respective scr RNA control samples.

Article Snippet: Additional OSMI1 compound was purchased from Sigma Aldrich (SML1621).

Techniques: Inhibition, MTS Assay, TUNEL Assay, Standard Deviation

Journal: Scientific Reports

Article Title: O-GlcNAc Transferase Inhibition Differentially Affects Breast Cancer Subtypes

doi: 10.1038/s41598-019-42153-6

Figure Lengend Snippet: Progesterone receptor inhibitor sensitizes receptor-positive BC cells to OGT inhibition. ( A ) Relative viability in MCF7 after 72 hours of treatment with Tamoxifen (Tam), OSMI1, or a combination. MTS assay. Error bars - SEM, n = 3. *p < 0.05; **p < 0.005, unpaired t-test, comparing viability after the combination treatment to the viability after treatment with respective dose of tamoxifen. ( B , C ) Viability in MCF7 and T47D cells following 72 hours of treatment with Mifepristone (MF), OSMI1 or a combination, MTS assay. Error bars - SEM, n = 3. *p < 0.05; **p < 0.005; ***p < 0.001 unpaired t-test, comparing viability after the combination-treatment to the viability after treatment with respective dose of MF. ( D ) Annexin V apoptosis assay in MCF7 following 72 hours of treatment with MF, OSMI1 or a combination. FITC-/PI- population represents live cells; FITC+ cells are in early apoptosis; FITC+/PI+ cells are in late apoptosis; PI+ cells are necrotic. Error bars - standard deviation, n = 2. ( E ) Total length and cleaved PARP and γ-H2AX in MCF7 after 72 hours of treatment with MF, OSMI1, or a combination. Representative of two separate experiments. ( F ) Relative γ-H2AX protein expression in MCF7 following 72 hours of treatment with MF, OSMI1 or a combination. n = 3, error bars - SEM; unpaired t-test, comparing expression after the combination treatments to the expression after respective concentration of MF and OSMI1; *p < 0.05.

Article Snippet: Additional OSMI1 compound was purchased from Sigma Aldrich (SML1621).

Techniques: Inhibition, MTS Assay, Apoptosis Assay, Standard Deviation, Expressing, Concentration Assay

Journal: Scientific Reports

Article Title: O-GlcNAc Transferase Inhibition Differentially Affects Breast Cancer Subtypes

doi: 10.1038/s41598-019-42153-6

Figure Lengend Snippet: OGT inhibition differentially affects proteome and phospho-proteome in MCF7 and MDA-MB-231. ( A ) Proteins whose expression changed by 28% or more compared to the control samples in MDA-MB-231 following 24 hours of treatment with 20 µM OSMI1, were selected. Corresponding proteins in MCF7 are shown in the same figure. RPPA array, n = 3, error bars - SEM. *p < 0.05; **p < 0.01, unpaired t-test. Significance indicates comparison of the relative protein expression changes in MDA-MB-231 and MCF7. ( B ) Total O-GlcNAcylation and phospho-PS6 (S235-S326) in MDA-MB-231 and MCF7 following treatment with 20 µM OSMI1 for 24 hours. ( C ) Phospho-PS6 (S235-S236) in MDA-MB-231 after treatment with 20 µM OSMI1 for 9, 16, 24, 32, 40 and 48 hours. ( D ) Viability in MCF7 and MDA-MB-231 following 72 hours of treatment with Everolimus (Ev), OSMI1 and the combination. MTS assay, error bars - SEM, n = 3. *p < 0.05, unpaired t-test. ( E , F ) Cell cycle distribution in MCF7 ( E ) and MDA-MB-231 ( F ), after 24 hours of treatment with 20 µM OSMI1. n = 3, error bars - SEM. *p < 0.05; **p < 0.005, unpaired t-test.

Article Snippet: Additional OSMI1 compound was purchased from Sigma Aldrich (SML1621).

Techniques: Inhibition, Expressing, MTS Assay

Journal: Scientific Reports

Article Title: O-GlcNAc Transferase Inhibition Differentially Affects Breast Cancer Subtypes

doi: 10.1038/s41598-019-42153-6

Figure Lengend Snippet: HES1 is selectively regulated by OGT in TNBC cells. ( A ) Relative protein HES1 expression following 24 hours of treatment with 20 µM OSMI1 (upper panel, error bars: SEM, n = 4; **p < 0.005, unpaired t-test) or 72 hours after a transient OGT knock-down (lower panel, error bars: St. dev, n = 2). ( B ) HES1 in MDA-MB-231 and MCF7 after 24 hours of treatment with 20 µM OSMI1. ( C ) HES1 protein expression in MDA-MB-231 following 2 hours of treatment with OSMI1, proteasome inhibitor MG-132, DMSO or combinations. Error bars - SEM, n = 3; *p < 0.05, unpaired t-test.

Article Snippet: Additional OSMI1 compound was purchased from Sigma Aldrich (SML1621).

Techniques: Expressing