Journal:

Article Title: Subtilosin Production by Two Bacillus subtilis Subspecies and Variance of the sbo-alb Cluster

doi: 10.1128/AEM.70.4.2349-2353.2004

Figure Lengend Snippet: Strains used

Article Snippet: Antibiotics were used at the following concentrations: 100 μg of ampicillin per ml and 20 μg of chloramphenicol per ml for E. coli and 5 μg of chloramphenicol per ml, 10 μg of kanamycin per ml, and 100 μg of spectinomycin per ml for B. subtilis. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Strain Other designations, description, and/or relevant genotype a Reference Bacillus subtilis strains ATCC 6633 CCM 1999, DSM 347, IAM 1069, NCIB 8054, NCTC 10400 6 168 Used for genome sequencing; DSM 402, NCIB 10106, B. subtilis subsp. subtilis 21 168 Δ sbo sbo :: cm (Cm r ) This study DSM 618 Test strain for detection of antibiotics in meat DSM 1088 IFO 13169, Bacillus natto DSM 2109 ATCC 11774, NCTC 8236 DSM 2277 ATCC 51189, CIP 103406, IAM 1633, NCIB 8649, NCTC 10073, B. atrophaeus 11 DSM 6405 Mutant of B. subtilis W23, B. subtilis subsp. spizizenii 21 60015 Marburg strain 10 10 T Type strain; ATCC 6051 T , CCM 2216 T , IAM 12118 T , IFO 13719 T , NCIB 3610 T , NCTC 3610 T ATCC 6633 16 Δ spaS spaS :: cm (Cm r ) Δ spaS spaS :: cm :: spec (Spec r ) This study Δ sbo sbo :: cm (Cm r ) This study Δ spaS Δ sbo spaS :: cm :: spec (Spec r ), sbo :: cm (Cm r ) This study Micrococcus luteus ATCC 9341 Test strain for antimicrobial activity Escherichia coli DH5α supE44 lacU169 [Φ80 lacZ M15] hsdR17 recA1 endA1 gyrA96 thi-1 relA1 23 Open in a separate window a ATCC, American Type Culture Collection; CCM, Czech Collection of Microorganisms; CIP, Collection de l'Institut Pasteur; DSM, German Collection of Microorganisms ( http://www.dsmz.de ); IAM, Institute of Applied Microbiology; IFO, Institute for Fermentation; NCIB, National Collection of Industrial Bacteria; NCTC, National Collection of Type Cultures.

Techniques: Sequencing, Mutagenesis, Activity Assay

Journal: Cell host & microbe

Article Title: SARS-CoV-2 Omicron sublineages show comparable cell entry but differential neutralization by therapeutic antibodies.

doi: 10.1016/j.chom.2022.04.017

Figure Lengend Snippet: Figure 1. S proteins of Omicron sublineages do not exhibit major differences in ACE2 usage or ability to drive cell-cell and virus-cell fusion (A) Schematic overview of the SARS-CoV-2 spike (S) protein domain structure (left) and summary of the mutations found in the different Omicron sublineages (right; numbering is according to the S protein of SARS-CoV-2 B.1). S protein residues that are identical between the S proteins of some Omicron sublineages and B.1 are marked in green, whereas mutated residues are highlighted in red (note: the BA.1 S protein harbors an insertion between amino acid residues 214 and 215). Further, mutations found in all Omicron sublineages are indicated by a circle. Abbreviations: NTD, N-terminal domain; RBD, receptor-binding domain; TD, transmembrane domain; S1/S2 and S2’, cleavage sites in the S protein. (B) S-protein-driven cell entry. We added particles bearing the indicated S proteins (or no S protein) to the indicated cell lines and analyzed cell entry by measuring the activity of virus-encoded firefly luciferase in cell lysates 16–18 h after inoculation. Presented are the average (mean) data from 6–12 biological replicates (each conducted with four technical replicates) in which cell entry was normalized against B.1 (set as 1). Error bars show the SEM. Statistical significance was assessed by two-tailed Student’s t tests (p > 0.05, not significant [ns]; p % 0.05, *; p % 0.01, **; p % 0.001, ***). Please also see Figure S1. (C) ACE2 binding efficiency. 293T cells transiently expressing the indicated S proteins (or no S protein) where first incubated with soluble ACE2 fused to the Fc portion of human immunoglobulin G (solACE2-Fc) and subsequently incubated with an Fc-specific AlexaFluor-488-coupled secondary antibody; then, solACE2- Fc binding was analyzed by flow cytometry. Presented are the average (mean) data from six biological replicates (each conducted with single samples) in which ACE2 binding was normalized against B.1 (set as 1). Error bars show the SEM. Statistical significance was assessed by two-tailed Student’s t tests (p > 0.05, not significant [ns]; p % 0.05, *; p % 0.01, **).

Article Snippet: 293T (human, female, kidney; ACC-635, DSMZ; RRID: CVCL_0063), Vero (African green monkey, female, kidney; CRL-1587, ATCC; RRID: CVCL_0603, kindly provided by Andrea Maisner), Huh-7 cells (human, male, liver; JCRB Cat# JCRB0403; RRID: CVCL_0336, e3 Cell Host & Microbe 30, 1103–1111.e1–e6, August 10, 2022 kindly provided by Thomas Pietschmann), BHK-21 (Syrian hamster, male, kidney; CCL-10, ATCC; RRID: CVCL_1915, kindly provided by Georg Herrler) were maintained in Dulbecco’s modified Eagle medium (DMEM, PAN-Biotech).

Techniques: Virus, Binding Assay, Activity Assay, Luciferase, Two Tailed Test, Expressing, Incubation, Cytometry

Journal: Nature Communications

Article Title: High-throughput identification of functional regulatory SNPs in systemic lupus erythematosus

doi: 10.1038/s41467-024-50710-5

Figure Lengend Snippet: A Flow chart illustrating the identification of functional candidate SNPs from SLE GWAS data. B SNP-seq: To generate the SNP-seq construct (top), a 31 bp sequence centered on the SNP is positioned between two type IIS restriction enzyme (IIS RE) binding sites. SNPs that fail to bind regulatory proteins such as transcription factors (TF) are negatively selected (bottom), allowing enrichment of protected constructs by PCR. The whole construct can be amplified using primers as per Supplementary Data . Bio biotin. Figure 1B was created with BioRender.com released under a Creative Commons Attribution-NonCommerical-NoDerivs 4.0 International license. C The experimental procedure for SNP-seq; NE nuclear extract, Bio biotin. D Spearman’s correlation with a two-tailed test of SNP-seq allele counts normalized to control between PBMC and BL2 samples (ρ = 0.89, P < 1 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\times$$\end{document} × 10 −15 ). E We selected 248 SNPs that passed next-generation sequencing quality control (NGS-QC) and demonstrated progressive allele-specific protection (Suppl. Fig. 1C). Source data are provided as a Source Data file.

Article Snippet: Human B cell line BL2 was from DSMZ (catalog number ACC 625) and human B cell line Daudi was kindly provided by Dr. Baglaenko Yuriy.

Techniques: Functional Assay, Construct, Sequencing, Binding Assay, Amplification, Two Tailed Test, Control, Next-Generation Sequencing

Journal: Nature Communications

Article Title: High-throughput identification of functional regulatory SNPs in systemic lupus erythematosus

doi: 10.1038/s41467-024-50710-5

Figure Lengend Snippet: A Seven biallelic and two triallelic (rs2297550 and rs936394) SNPs showed consistent allele-imbalanced binding for nuclear by EMSA. Allele-specific gel shift/binding marked with red circles. Experiments were repeated three times using distinct nuclear extracts. Representative blots from PBMCs are shown here; BL2 data and negative SNPs are provided in Supplementary Fig. and Supplementary Data . B Luciferase reporter assay between the reference (red) and alternative (blue) alleles of nine candidate regulatory SNPs from A in Daudi B cells (mean \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm$$\end{document} ± s.d, n = 8 biological replicates, Mann–Whitney two-tailed U -test with two-stage step-up correction for multiple hypothesis testing: rs2297550 p = 0.00047, rs906868 p = 0.0013, rs936394 p = 0.00047, rs9907966 p = 0.0071, rs13213604 p = 0.0085). C Diagram displaying the position of 5 SNPs that showed consistent significant differences between alleles through EMSA and luciferase reporter assay; rs2297550 in promoter of IKBKE , rs906868 in intergenic region between YPEL5 and LBH , rs936394 in intron of WBP2 , rs9907966 in intron of IKZF3 , rs13213604 in intron of BLTP3A . The unit of chromosome position is in kilobases (Kb). A diagram of gene features including exon, intron, and UTR was generated using GSDS 2.0 ( http://gsds.gao-lab.org/index.php ). Source data are provided as a Source Data file.

Article Snippet: Human B cell line BL2 was from DSMZ (catalog number ACC 625) and human B cell line Daudi was kindly provided by Dr. Baglaenko Yuriy.

Techniques: Binding Assay, Gel Shift, Luciferase, Reporter Assay, MANN-WHITNEY, Two Tailed Test, Generated

Journal: Nature Communications

Article Title: Circular EZH2-encoded EZH2-92aa mediates immune evasion in glioblastoma via inhibition of surface NKG2D ligands

doi: 10.1038/s41467-022-32311-2

Figure Lengend Snippet: a Illustration of the in situ GBM model in immunocompromised mice and treatment with NK cells. b Kaplan–Meier analysis of mice intracranially implanted with MES28 and GSC23 GSCs with stable KD of circEZH2 or control cells and treated with PBS or NK cells ( n = 5 per group). Log-rank test. ** P < 0.01. MST, median survival time. Scale bar, 1 mm. Sh-circEZH2+NK vs scramble, MES28 P = 0.0023, GSC23 P = 0.0017; sh-circEZH2+NK vs scramble+NK, MES28 P = 0.0026, GSC23 P = 0.0017; sh-circEZH2+NK vs sh-circEZH2, MES28 P = 0.0025, GSC23 P = 0.0025. c Representative H&E-stained brain slices from mice with indicated treatment in ( b ). Scale bar, 1 mm. d , e Representative histogram ( d ) and quantification ( e ) of the expression of the indicated molecules in NK cells isolated from the abovementioned GBM model ( n = 8 per group). scr vs sh, MES28, CD107a P = 7.37e−07, GZMB P = 5.92e−04, Perforin P = 1.97e−05, IFN-γ P = 1.17e−06, TNF-α P = 1.36e−05; GSC23, CD107a P = 2.21e−05, GZMB P = 3.96e−04, Perforin P = 1.09e−04, IFN-γ P = 0.0028, TNF-α P = 0.0040. f Illustration of the in situ GBM model in C57BL/6 mice and treatment with NK cells and the anti-PD1 antibody. g Kaplan–Meier analysis of mice intracranially implanted with GL261 cells with stable circEZH2 KD or scrambled control cells and treated with PBS, NK cells or NK cells combined with an anti-PD1 antibody ( n = 5 per group). The log-rank test was performed between the indicated groups. MST, median survival time. h Representative images of H&E-stained brain slices from mice with indicated treatment in ( g ). Scale bar, 1 mm. i Frequencies of NK or CD8 + T cells positive with the indicated molecules isolated from the tumour mass from the abovementioned GL261 model ( n = 5 per group). For NK cell panel, scr+NK vs sh+NK, CD107a P = 0.0085, GZMB P = 0.0020, IFN-γ P = 0.0006; sh+NK vs sh+NK + anti-PD1, CD107a P = 0.0102, GZMB P = 0.0007, IFN-γ P = 5.61e−05; for CD8 T cell panel, scr+NK vs scr+NK + anti-PD1, GZMB P = 0.0041, IFN-γ P = 0.0035; sh+NK vs sh+NK + anti-PD1, GZMB P = 0.0224, IFN-γ P = 0.0006. The data are presented as the mean ± SD values. Unpaired two-tailed Student’s t test was used to determine the significance of differences between the indicated groups. ns, nonsignificant, * P < 0.05; ** P < 0.01; *** P < 0.001. Source data are provided as a Source data file.

Article Snippet: 293T cells (ATCC, CRL-3216) and GL261 (DSMZ, ACC 802) were cultured in DMEM (Gibco) supplemented with 10% FBS according to standard protocols.

Techniques: In Situ, Control, Staining, Expressing, Isolation, Two Tailed Test

Journal: Scientific Reports

Article Title: A common theme in extracellular fluids of beetles: extracellular superoxide dismutases crucial for balancing ROS in response to microbial challenge

doi: 10.1038/srep24082

Figure Lengend Snippet: Thirty larvae were injected each with 200 ng of PcSOD3.1 -, PcSOD3.2-, PcSOD1- dsRNA or gfp -dsRNA. Five days after this initial injection, larvae were challenged with microbes. The probability of survival is depicted as Kaplan-Meier curves by applying Gehan-Breslow statistic test. The weight of freshly emerged pupa was determined after RNAi silencing and infection of larvae. ( A ) Survival upon fungal challenge. After emptying of the defence reservoirs, the larvae of each group were submerged at the same time in the same solution of M. anisopliae 1*10 6 conidia/ml. Overall level of significance: p = 0.014 (Gehan-Breslow test); p = 0.04 (Log Rank (Mantel-Cox)). Pairwise multiple comparison procedures for gfp vs. Pc SOD3.1(−): p = 0.04 (Holm-Sidak method). Pupal weight, level of significance: p = 0.023* (two-tailed t-test). ( B ) Survival after injection of 1*10 6 E. coli -cells/200 nl. ( C ) Survival after injection of 1*10 6 M. luteus -cells/200 nl.

Article Snippet: Metarhizium anisopliae (DSM-1490, DSMZ, Braunschweig, Germany) was maintained on potato-dextrose-agar (Sigma, 2% glucose), and mycelia were used for subsequent propagation.

Techniques: Injection, Infection, Comparison, Two Tailed Test

Journal: Cell host & microbe

Article Title: An early-life microbiota metabolite protects against obesity by regulating intestinal lipid metabolism.

doi: 10.1016/j.chom.2023.09.002

Figure Lengend Snippet: Figure 2. Ligilactobacillus murinus protects against adiposity during early-life consumption of an HF diet (A) Heatmap showing differential abundance of the 5 most abundant families in the small intestine microbiota of mice after 5 weeks of treatment. (B) Relative abundance of the genus Lactobacillus in the small intestine microbiota as determined by 16S rRNA sequencing. (C) The abundance of L. murinus in the small intestine as determined by qPCR. (D and E) (D) Weight gain and (E) abdominal fat of mice given an HF diet and LDP alone or gavaged with a penicillin resistant strain of L. murinus (L. murinus PenR) after 5 weeks. (F) A schematic representation of the gnotobiotic experiment. (G) Lactobacillus abundance (colony-forming unit [CFU]/gram) in small intestine (SI) after 5 weeks on an HF. Dotted line indicates limit of detection for Lacto- bacillus species in this experiment. (H and I) (H) Weight gain and (I) abdominal fat (g) of mice after 5 weeks. Each dot represents one animal. Bars represent geometric mean. (A) n = 5 mice/group; (B) n = 9 mice/group; (C) n = 7–8 mice/group; (D and E) n = 12 mice/group; (G and H) n = 8 mice/group. Data representative of two in- dependent cohorts. *p < 0.05; **p < 0.01; ***p < 0.005; ****p < 0.001 using an unpaired two-tailed Student’s t test.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-PPAR-g Santa Cruz Biotechnology Cat #: sc-7273, RRID:AB_628115 Goat anti-mouse IgG Alexa Fluor 488 Thermo Fisher Scientific Cat #: A-11001, RRID:AB_2534069 Bacterial and virus strains Ligilactobacillus murinus This study CS010 Muribaculum intestinale DSMZ YL27 Clostridium clostridioforme DSMZ YL32 Clostridium sporogenes DSMZ B-CC-163-3B Streptococcus daniellae ERD01G DSMZ ERD01G Staphylococcus xylosus 33-ERD13C DSMZ 33-ERD13C Escherichia coli Mt1B1 DSMZ Mt1B1 Ligilactobacillus murinus PenR This study CS052 Ligilactobacillus murinus DSMZ M-6244-3B Lacticaseibacillus rhamnosus ATCC Cat #: 53103 Lactobacillus acidophilus ATCC Scav; Cat #: 4356 Bifidobacterium longum subsp animalis DSMZ YL2 Chemicals, peptides, and recombinant proteins Difco Lactobacilli MRS agar Becton Dickinson Cat #: 288210 Difco Lactobacilli MRS broth Becton Dickinson Cat #: 288130 Penicillin V potassium salt Sigma Cat #: P1382 D-(+)-3-phenyllactic acid Sigma Cat #: 376906 Difco LB broth Becton Dickinson Cat #: 244620 Difco Brain Heart Infustion agar Becton Dickinson Cat #: 241830 Bacto Yeast Extract Becton Dickinson Cat #: 212750 Bacto Tryptic soy broth Becton Dickinson Cat #: 211825 Dipotassium hydrogen phosphate Fisher Chemicals Cat #: P288 Hemin Sigma Cat #: 51280 D(+)-Glucose Acros Organics Cat #: 41095 Sodium carbonate Fisher Chemicals Cat #: 5263 (L) – cysteine Sigma Cat #: 168149 Menadione Sigma Cat #: 47775 Heat-Inactivated Fetal Bovine Serum Gibco Cat #: 16140-071 RPMI 1640 Medium Gibco Cat #: 11875093 Recombinant Mouse IFN-gamma Protein R&D Systems Cat #: 485-MI-100 Penicillin-Streptomycin Gibco Cat #: 15140122 ITS Liquid Media Supplement Sigma Cat #: I3146 Minimal Essential Media Gibco Cat #: 11095-080 Glutamax Gibco Cat #: 35050-061 Nonessential amino acids Gibco Cat #: 11140-050 Sodium pyruvate Gibco Cat #: 11360070 Trypsin-EDTA (0.05%) Gibco Cat #: 25300054 Primocin InvivoGen Cat #: ant-pm-1 Dithiothreitol Acros Organics Cat #: 16568 EDTA Sigma Cat #: E5134 EGTA Sigma Cat #: E3889 (Continued on next page) e1 Cell Host & Microbe 31, 1604–1619.e1–e10, October 11, 2023

Techniques: Sequencing, Two Tailed Test

Journal: Journal for Immunotherapy of Cancer

Article Title: Bone marrow MSC from pediatric patients with B-ALL highly immunosuppress T-cell responses but do not compromise CD19-CAR T-cell activity

doi: 10.1136/jitc-2020-001419

Figure Lengend Snippet: In vitro cytotoxicity of CD19-CAR T-cells against B-ALL cell lines in the absence/presence of BM-MSC. (A) Achematic of the experimental design for assessing the cytotoxicity of CD19-CAR T-cells against NALM6 or SEM B-ALL cell lines in the absence or presence of BM-MSC. (B, C) Absolute counts of alive target cells NALM6 (B) and SEM (C) measured by FACS after 2 and 6 days at 1:1 E:T ratio in the absence/presence of BM-MSC. Data are shown as mean±SEM. ****P<0.0001; one-way ANOVA test with Tukey’s post hoc test. HD BM-MSC n=3 and B-ALL BM-MSC n=5. ANOVA, analysis of variance; B-ALL, B-cell acute lymphoblastic leukemia; BM-MSC, bone marrow-mesenchymal stem/stromal cells; E:T, Effector:Target; HD, healthy donors.

Article Snippet: The B-ALL cell lines SEM and NALM6 were obtained from DSMZ cell line bank, Braunschweig, Germany.

Techniques: In Vitro

Journal: Journal for Immunotherapy of Cancer

Article Title: Bone marrow MSC from pediatric patients with B-ALL highly immunosuppress T-cell responses but do not compromise CD19-CAR T-cell activity

doi: 10.1136/jitc-2020-001419

Figure Lengend Snippet: In vivo cytotoxicity of CD19-CAR T-cells in an NALM6 mouse xenograft model in the absence/presence of BM-MSC. (A) Schematic of the transplantation of B-ALL cells and BM-MSC followed by CD19-CAR T-cell infusion after 4 days to assess in vivo the influence of BM-MSC on CD19-CAR T-cell activity. (B) Leukemic burden monitored weekly (until sacrifice of control mice) by BLI. (C) Total RADIANCE quantification (p/sec/cm 2 /sr) of mice shown in B. (D) Leukemic burden analyzed by FACS in PB and BM at the time control mice were sacrificed (day 15). (E) Representative FACS dot plots identifying target cells (red) and T-cell persistence (blue) in BM at the time control mice were sacrificed. (F) Long-term (35 days) BLI follow-up of CD19-CAR T-cell-treated mice in the absence/presence of BM-MSC. (G) Total RADIANCE quantification (p/sec/cm 2 /sr) of mice shown in F). Each dot represents a mouse. Data are shown as mean±SEM. *P<0.05, ***p<0.001, ****p<0.0001; a paired two-tailed t-test was used when two experimental groups were compared, and one-way ANOVA test with Tukey’s post hoc test was used when three experimental groups were compared. ANOVA, analysis of variance; B-ALL, B-cell acute lymphoblastic leukemia; BM-MSC, bone marrow-mesenchymal stem/stromal cells; CAR, chimeric antigen receptor; E:T, Effector:Target; PB, peripheral blood.

Article Snippet: The B-ALL cell lines SEM and NALM6 were obtained from DSMZ cell line bank, Braunschweig, Germany.

Techniques: In Vivo, Transplantation Assay, Activity Assay, Control, Two Tailed Test

Journal: Nature communications

Article Title: Distinctive phenotypes and functions of innate lymphoid cells in human decidua during early pregnancy.

doi: 10.1038/s41467-019-14123-z

Figure Lengend Snippet: Fig. 6 dNK responses triggered by activating KIR. a Representative XCL1 and CD107a staining by CyTOF in lineage negative (Lin−) CD56+ decidual cells co-expressing 1–3 Killer-cell immunoglobulin-like receptors (KIR) following a 6 h co-culture with K562 (n = 10). b Representative XCL1 and CD107a staining by mass cytometry in Lin-CD56+ cells co-expressing 1–3 KIR following 4 h stimulation by PMA plus ionomycin (n = 8). c Representative XCL1 staining by flow cytometry in Lin-CD56+KIR2DS4+ decidual cells co-expressing 1–3 additional KIRs following activation via P815 cells coated with anti-KIR2DS4 (n = 6). d Correlation of frequency of XCL1+Lin-CD56+ decidual cells and mean side scatter for the same subset (n = 20). Two-tailed p-value calculated for Pearson correlation coefficients. Two-tailed one-way ANOVA of matched data points with Tukey correction and 95% confidence level was used. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.

Article Snippet: The murine cell line P815 and human cell line K562 were purchased from DSMZ in Germany.

Techniques: Staining, Expressing, Co-Culture Assay, Mass Cytometry, Cytometry, Activation Assay, Two Tailed Test

Journal: Pharmaceutics

Article Title: Combating Acute Myeloid Leukemia via Sphingosine Kinase 1 Inhibitor-Nanomedicine Combination Therapy with Cytarabine or Venetoclax

doi: 10.3390/pharmaceutics16020209

Figure Lengend Snippet: MP-A08-loaded liposomes chemosensitised human AML cell line MV4-11 to cytarabine. Cell viability assays of cytarabine in combination with MP-A08-loaded liposomes were assessed in MV4-11 cell line. MV4-11 was treated with blank liposomes, MP-A08-loaded liposomes (0.125 µM), and cytarabine [( a ) 0.1 µM, ( b ) 0.25 µM, ( c ) 0.5 µM] as a single agent or in combination with MP-A08-loaded liposomes for 48 h. Cell viability was assessed using annexin V and PI staining by flow cytometry. Each experiment contained three replicates for every drug combination. Error bars indicate the mean ± range of 3 replicates of representative experiment of n = 3 experiments. Synergism was determined by the Chou–Talalay Combination Index (CI) using CompuSyn software. CI values are shown for distinct dose combinations of MP-A08-loaded liposomes and cytarabine. p values were calculated using a two-tailed Student’s t -test, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: AML Cell Lines: the human AML cell lines used in this study were OCI-AML3, obtained from the Leibniz Institute DSMZ in Braunschweig, Germany, and MV4-11 cells, acquired from the American Type Culture Collection (ATCC) in Manassas, VA, USA.

Techniques: Liposomes, Staining, Flow Cytometry, Software, Two Tailed Test

Journal: Pharmaceutics

Article Title: Combating Acute Myeloid Leukemia via Sphingosine Kinase 1 Inhibitor-Nanomedicine Combination Therapy with Cytarabine or Venetoclax

doi: 10.3390/pharmaceutics16020209

Figure Lengend Snippet: MP-A08-loaded liposomes chemosensitised human AML cell line OCI-AML3 to cytarabine. Cell viability assays of cytarabine in combination with MP-A08-loaded liposomes were assessed in OCI-AML3 cell line. OCI-AML3 was treated with blank liposomes, MP-A08-loaded liposomes (0.125 µM), and cytarabine [( a ) 0.1 µM, ( b ) 0.25 µM, ( c ) 0.5 µM] as a single agent or in combination with MP-A08-loaded liposomes for 48 h. Cell viability was assessed using annexin V and PI staining by flow cytometry. Each experiment contained three replicates for every drug combination. Error bars indicate the mean ± range of 3 replicates of representative experiment of n = 3 experiments. Synergism was determined by the Chou–Talalay Combination Index (CI) using CompuSyn software. CI values are shown for distinct dose combinations of MP-A08-loaded liposomes and cytarabine. p values were calculated using a two-tailed Student’s t -test, * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: AML Cell Lines: the human AML cell lines used in this study were OCI-AML3, obtained from the Leibniz Institute DSMZ in Braunschweig, Germany, and MV4-11 cells, acquired from the American Type Culture Collection (ATCC) in Manassas, VA, USA.

Techniques: Liposomes, Staining, Flow Cytometry, Software, Two Tailed Test

Journal: Pharmaceutics

Article Title: Combating Acute Myeloid Leukemia via Sphingosine Kinase 1 Inhibitor-Nanomedicine Combination Therapy with Cytarabine or Venetoclax

doi: 10.3390/pharmaceutics16020209

Figure Lengend Snippet: MP-A08-loaded liposomes potentiated venetoclax anti-leukemic activity in MV4-11 AML cell line. Cell viability assays of venetoclax in combination with MP-A08-loaded liposomes were assessed. Representative data of n = 3 experiments, each containing three replicates for every drug combination (error bars indicating the mean ± range of 3 replicates). MV4-11 human AML cell lines were treated with blank liposomes, MP-A08 liposomes (0.125 µM), and venetoclax [( a ) 0.5 nM, ( b ) 1 nM, ( c ) 2.5 nM] as a single agent or in combination for 48 h. Cell viability was assessed using annexin V and PI staining via flow cytometry. Synergism was determined by the Chou–Talalay Combination Index (CI) using CompuSyn software. CI values are shown for distinct dose combinations of MP-A08-loaded liposomes and venetoclax. p values were calculated using a two-tailed Student’s t -test, * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: AML Cell Lines: the human AML cell lines used in this study were OCI-AML3, obtained from the Leibniz Institute DSMZ in Braunschweig, Germany, and MV4-11 cells, acquired from the American Type Culture Collection (ATCC) in Manassas, VA, USA.

Techniques: Liposomes, Activity Assay, Staining, Flow Cytometry, Software, Two Tailed Test

Journal: Pharmaceutics

Article Title: Combating Acute Myeloid Leukemia via Sphingosine Kinase 1 Inhibitor-Nanomedicine Combination Therapy with Cytarabine or Venetoclax

doi: 10.3390/pharmaceutics16020209

Figure Lengend Snippet: MP-A08-loaded liposomes enhanced venetoclax anti-leukemic activity in venetoclax-resistant AML cell line OCI-AML3. Cell viability assays of venetoclax in combination with MP-A08-loaded liposomes were assessed. Representative data of n = 3 experiments, each containing three replicates for every drug combination (error bars indicating the mean ± range of 3 replicates). Venetoclax-resistant human AML OCI-AML3 cells were treated with blank liposomes, MP-A08-loaded liposomes (0.125 µM), or venetoclax [( a ) 0.05 µM, ( b ) 0.1 µM, ( c ) 0.25 µM] as a single agent or in combination for 48 h. Cell viability was assessed using annexin V and PI staining by flow cytometry. Synergism was determined by the Chou–Talalay Combination Index (CI) using CompuSyn software. CI values are shown for distinct dose combinations of MP-A08-loaded liposomes and venetoclax. p values were calculated using a two-tailed Student’s t -test, ** p < 0.01, *** p < 0.001 and **** p < 0.001.

Article Snippet: AML Cell Lines: the human AML cell lines used in this study were OCI-AML3, obtained from the Leibniz Institute DSMZ in Braunschweig, Germany, and MV4-11 cells, acquired from the American Type Culture Collection (ATCC) in Manassas, VA, USA.

Techniques: Liposomes, Activity Assay, Staining, Flow Cytometry, Software, Two Tailed Test

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

Article Title: Inactivation of PRIM1 Function Sensitizes Cancer Cells to ATR and CHK1 Inhibitors 1

doi: 10.1016/j.neo.2018.08.009

Figure Lengend Snippet: siPRIM1-mediated sensitization to ATR and CHK1 inhibitors in a panel of different cancer cell lines. (A) Knockdown efficiency of PRIM1 via siRNA was confirmed by immunoblotting 96 hours after transfection. β-Actin was used as loading control. (B) Effects of ATR and (C) CHK1 inhibitors on the proliferation of PRIM1 -depleted SW480, RKO, and PaTu 8988t cells as compared to control and mock-transfected cells measured 120 hours after drug treatment. Error bars represent ±SEM of at least three independent experiments with each data point reflecting triplicate wells. Asterisks mark statistical significance using a two-tailed, unpaired Student's t test (* P < .05, ** P < .01, *** P < .001). Immunoblots were performed independently at least three times, and representative results are shown.

Article Snippet: The human colorectal cancer (CRC) cell lines DLD-1, SW480, and RKO were purchased from the Leibniz Institut DSMZ (Braunschweig, Germany) or the American Type Culture Collection (LGC Standards, Wesel, Germany), respectively.

Techniques: Knockdown, Western Blot, Transfection, Control, Two Tailed Test