methylcellulose methocult h4534 classic  (STEMCELL Technologies Inc)

Journal: Nature

Article Title: Perturbing LSD1 and WNT rewires transcription to synergistically induce AML differentiation

doi: 10.1038/s41586-025-08915-1

Figure Lengend Snippet: a , Mouse AML cells treated with 50 nM GSK–LSD1 and different concentrations of LY2090314 for 5 days; cell growth was determined by the Cell-Titer-Glo assay. Data are presented as mean ± s.d. from two biological independent experiments. BMDM, bone marrow-derived macrophage; RLU, relative light unit. b , Quantification of colonies formed by normal mouse LSK cells treated with the indicated inhibitors. Data are presented as mean ± s.d. from three biological independent experiments. P values were determined using two-way ANOVA. NS, not significant. c , Flow-cytometric quantification of CD11b and Gr-1 expression in the cells harvested from methylcellulose in panel b . d , Analysis of cell viability in THP-1 cells treated with the indicated inhibitors for 5 days by the Cell-Titer-Glo assay. Data are presented as mean ± s.d. from three biological independent experiments. P values were determined using two-way ANOVA. e , Analysis of CD11b mRNA relative level in THP-1 cells treated with the indicated inhibitors for 5 days. Values were normalized against GAPDH . Data are presented as mean ± s.d. from three biological independent experiments. P values were determined using two-way ANOVA. f , Quantification of colonies formed by THP-1 cells treated with the indicated inhibitors. Data are presented as mean ± s.d. from three biological independent experiments. P values were determined using two-way ANOVA. g , h , Tumour burden ( g ) and Kaplan–Meier survival curves ( h ) of mice treated with vehicle ( n = 15), GSK–LSD1 ( n = 9), LY2090314 ( n = 11) or a combination of both inhibitors ( n = 20) in a syngeneic model of a HOXA9–MEIS1-driven AML model. P values were determined using the log-rank test. The error bars represent mean ± s.e.m. ( g ). P values were determined using two-way ANOVA.

Article Snippet: Human cord blood CD34 + cells (200-0000, StemCell Technologies) were plated in methylcellulose (MethoCult H4534 Classic, StemCell Technologies).

Techniques: Glo Assay, Derivative Assay, Expressing

Journal: Oncotarget

Article Title: Pre-clinical evidence of PIM kinase inhibitor activity in BCR-ABL1 unmutated and mutated Philadelphia chromosome-positive (Ph+) leukemias

doi:

Figure Lengend Snippet: A. K562 cells were treated with control (DMSO), SGI-1776 (3 μmol/L) or imatinib (0.5 μmol/L) alone or in combination for two hours, and equal amounts of cell lysates were resolved by SDS-PAGE and immunoblotted with the indicated antibodies. The immunoblots with antibodies against the phosphorylated forms of the proteins or against the total proteins were from lysates from the same experiments analyzed in parallel by SDS-PAGE. B. KT1 cells were treated with control (DMSO), SGI-1776 (3 μmol/L) or imatinib (1 μmol/L) alone or in combination for two hours, and equal amounts of cell lysates were resolved by SDS-PAGE and immunoblotted with the indicated antibodies. The immunoblots with antibodies against the phosphorylated forms of the proteins or against the total proteins were from lysates from the same experiments analyzed in parallel by SDS-PAGE. C. K562 cells were plated in methylcellulose in the presence of control (DMSO), SGI-1776 (10 μmol/L) or imatinib (0.5 μmol/L) or in combination for approximately seven days. Leukemic progenitor colony formation (CFU-L) was assessed in clonogenic assays in methylcellulose. Data are expressed as a percentage of DMSO control-treated cells. Shown are the means + SE of six independent experiments. *** p < 0.001, **** p < 0.0001 using a paired t-test. D. Representative colonies treated with control (DMSO), SGI-1776, imatinib, and SGI-1776 plus imatinib, from the experiments described in panel C is shown (x 10 magnification). E. LAMA-84 cells were plated in methylcellulose in the presence of control (DMSO), SGI-1776 (3 μmol/L) or imatinib (0.5 μmol/L) alone or in combination. Leukemic progenitor colony formation (CFU-L) was assessed in clonogenic assays in methylcellulose. Data are expressed as a percentage of DMSO control-treated cells. Shown are the means + SE of five independent experiments. * p < 0.05, ** p < 0.01 using a paired t-test. F. K562 cells were treated with control (DMSO), SGI-1776 (3 μmol/L) or imatinib (0.5 μmol/L) alone or in combination for 24 hours. The percentage of apoptosis was determined by flow cytometry using Annexin V and DAPI stain. Shown are the means + SE of three independent experiments. * p < 0.05, ** p < 0.01 using a paired t-test G. KT1 cells were treated with control (DMSO), SGI-1776 (3 μmol/L) or imatinib (1 μmol/L) alone or in combination for 24 hours. The percentage of apoptosis was determined by flow cytometry using Annexin V and DAPI stain. Shown are the means + SE of three independent experiments. *** p < 0.001, **** p < 0.0001 using a paired t-test.

Article Snippet: To assess the effects of drugs on leukemic progenitor (CFU-L) colony formation, cells were then plated in methylcellulose (MethoCult™ H4534 Classic without EPO, Stem Cell Technologies).

Techniques: SDS Page, Western Blot, Flow Cytometry, Staining

Journal: Oncotarget

Article Title: Pre-clinical evidence of PIM kinase inhibitor activity in BCR-ABL1 unmutated and mutated Philadelphia chromosome-positive (Ph+) leukemias

doi:

Figure Lengend Snippet: A. Primary mononuclear cells derived from CML patient samples were plated in methylcellulose in the presence of control (DMSO) or SGI-1776 at the indicated concentrations for approximately fourteen days. Leukemic progenitor colony formation (CFU-GM) was assessed in clonogenic assays in methylcellulose. Data are expressed as a percentage of DMSO control-treated cells. Shown are the means + SE of five experiments performed with samples from five different patients. *** p < 0.001 using a paired t -test. B. Primary mononuclear cells derived from CML patient samples were plated in methylcellulose in presence of control (DMSO), SGI-1776 (0.5 μmol/L) or imatinib (0.5 μmol/L) alone or in combination for approximately fourteen days, as indicated. Leukemic progenitor colony formation (CFU-GM) was assessed in clonogenic assays in methylcellulose. Data are expressed as a percentage of DMSO control-treated cells. Shown are the means + SE of five experiments performed with samples from five different patients. * p < 0.05, ** p < 0.01, *** p < 0.001 using a paired t -test C. Primary mononuclear cells derived from CML patient samples were plated in methylcellulose in the presence of control (DMSO) or AZD-1208 at the indicated doses for approximately fourteen days. Leukemic progenitor colony formation (CFU-GM) was assessed in clonogenic assays in methylcellulose. Data are expressed as a percentage of DMSO control-treated cells. Shown are the means + SE of four experiments performed with samples from four different patients. ** p < 0.01 using a paired t -test.

Article Snippet: To assess the effects of drugs on leukemic progenitor (CFU-L) colony formation, cells were then plated in methylcellulose (MethoCult™ H4534 Classic without EPO, Stem Cell Technologies).

Techniques: Derivative Assay