Journal: Cell host & microbe

Article Title: An active HIV reservoir during ART is associated with maintenance of HIV-specific CD8 + T cell magnitude and short-lived differentiation status

doi: 10.1016/j.chom.2023.08.012

Figure Lengend Snippet:

Article Snippet: QuantiNova Probe PCR Kit , QIAGEN , Cat# 80224.

Techniques: Recombinant, Staining, One Step RT-PCR, Software, Northern Blot, Cytometry, In Vitro

Journal: PLoS ONE

Article Title: Blockade of p38 MAPK overcomes AML stem cell line KG1a resistance to 5-Fluorouridine and the impact on miRNA profiling

doi: 10.1371/journal.pone.0267855

Figure Lengend Snippet: (A) Representative cell scatter plots and histogram analysis of leukemic stem-like AML cell line KG1a after immuno-cyto-staining with FITC-labeled anti-CD34 antibody, with APC-labeled anti-CD38 antibody or with respective IgG2a-FITC and IgG2a-APC isotype controls confirming the phenotypic characteristic of the chemoresistance potential of stem cells defined as CD34 + /CD38 - . Bar graph showing the positive expression rates of KG1a cells based on CD34 and CD38 cell surface detection. (B) Transcriptomic profiling and analysis of differentially expressed genes in AML KG1a stem cells and HL60 cells. Using the Clariom™ D Human Transcriptomic array, selection of the top 100 differentially expressed genes. (C) KG1a and HL60 cell characterization based on selected transcriptomic profiling using the ClariomD™ array. (D) Highlight on the top differentially expressed drug resistance-related genes in HL60 and KG1a cells. (E, F) Bar graphs showing the percentage viability of the KG1a and HL60 cells treated with various concentrations (0.001–10 μM) of either Doxorubicin (D) or 5-Fluorouridine (E). After 72 h incubation, the cell viability was determined using CellTiter-Glo ® and expressed as percentage of the control, the non-treated cell viability, corresponding to 100%. The results are presented as mean ± SD and were repeated through three independent experiments. * p < 0.05, *** p < 0.001, and **** p < 0.0001 vs . control.

Article Snippet: The human AML stem cell line KG1a (#CCL-246.1) and Caucasian promyelocytic leukemia cell line HL60 (#CCL-240) were provided by the American Type Culture Collection (Manassas, VA, USA).

Techniques: Staining, Labeling, Expressing, Selection, Incubation, Control

Journal: PLoS ONE

Article Title: Blockade of p38 MAPK overcomes AML stem cell line KG1a resistance to 5-Fluorouridine and the impact on miRNA profiling

doi: 10.1371/journal.pone.0267855

Figure Lengend Snippet: The KG1a (A) and HL60 (B) cells were treated with 5 mM NAC, 20 μM IKKVII and 10 μM SB202190. After 72 h incubation, viable, (early and late) apoptotic, and necrotic status of the cells were determined using Apoptosis determination kit. Representative cell scatter plots indicating the percentage of cells determined at each status. Bar graphs showing the results presented as mean ± SD, based on three independent experiments. * p < 0.05, *** p < 0.001, and **** p < 0.0001 vs . control.

Article Snippet: The human AML stem cell line KG1a (#CCL-246.1) and Caucasian promyelocytic leukemia cell line HL60 (#CCL-240) were provided by the American Type Culture Collection (Manassas, VA, USA).

Techniques: Incubation, Control

Journal: PLoS ONE

Article Title: Blockade of p38 MAPK overcomes AML stem cell line KG1a resistance to 5-Fluorouridine and the impact on miRNA profiling

doi: 10.1371/journal.pone.0267855

Figure Lengend Snippet: The KG1a and HL60 cells were pre-treated with 10 μM SB202190 for 2 h incubation then followed by the cell treatment with 10 μM of FUrd. After 72 h incubation, the cell lysates were subjected to Western blot analysis. Representative Western blot showing the decrease of the p38 phosphorylation by SB202190, confirming the blockade of the p38 MAPK pathway. The bar graphs show the relative protein expression levels of phospho-p38 calculated as a ratio of total p38 expression (the loading control). The results are presented as mean ± SD of three independent experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs . control. Tubulin was used as a loading control.

Article Snippet: The human AML stem cell line KG1a (#CCL-246.1) and Caucasian promyelocytic leukemia cell line HL60 (#CCL-240) were provided by the American Type Culture Collection (Manassas, VA, USA).

Techniques: Incubation, Western Blot, Phospho-proteomics, Expressing, Control

Journal: PLoS ONE

Article Title: Blockade of p38 MAPK overcomes AML stem cell line KG1a resistance to 5-Fluorouridine and the impact on miRNA profiling

doi: 10.1371/journal.pone.0267855

Figure Lengend Snippet: Analysis of apoptosis status in KG1a cells in response to 24-h (A) and 72-h (B) treatment with FUrd after cell pretreatment with SB202190. The KG1a cells were non-treated, treated with pharmacological inhibitors for 2 h incubation prior to cell treatment with 10 μM of FUrd. After 24 h (A) and 72 h (B) incubation, the cells underwent FITC-labeled Annexin V/PE-labeled PI double-staining for apoptosis analysis using FACS. The numbers within the scatter plots represent the percentage of viable (lower left, Annexin V - /PI - ), early apoptotic (lower right, Annexin V + /PI - ), late apoptotic (upper right, Annexin V + /PI + ), and necrotic cells (upper left, Annexin V - /PI + ). The bar graphs present the percentage viable, early apoptosis, late apoptosis and necrosis of SB202190-pretreated KG1a cells in the presence or absence of 10 μM FUrd along with the non-treated (control) cells, based on at least three independent experiments. The results are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs . control.

Article Snippet: The human AML stem cell line KG1a (#CCL-246.1) and Caucasian promyelocytic leukemia cell line HL60 (#CCL-240) were provided by the American Type Culture Collection (Manassas, VA, USA).

Techniques: Incubation, Labeling, Double Staining, Control

Journal: PLoS ONE

Article Title: Blockade of p38 MAPK overcomes AML stem cell line KG1a resistance to 5-Fluorouridine and the impact on miRNA profiling

doi: 10.1371/journal.pone.0267855

Figure Lengend Snippet: Induction of poly-caspase activity and mitochondrial outer membrane permeabilization activity in the HL60 cells (A-C) and in the KG1a (D-F) in response to FUrd after pretreatment with SB202190. Both the HL60 and KG1a cells were non-treated, treated with SB202190 for 2 h incubation prior to cell treatment with 10 μM of FUrd. After 24 h incubation for the HL60 cells (A-C) and 72 h incubation for the KG1a cells (D-F), the cells underwent staining for assessment of poly-caspase activity (A, D) and of mitochondrial outer membrane polymerization (B, E) in apoptotic cells (indicated in green). The bar graphs present the percentage of the poly-caspase activity and of the mitochondrial outer membrane polymerization determined in SB202190-pretreated HL60 (C) and KG1a (F) cells in the presence or absence of FUrd along with the non-treated (control) cells. The results are presented as mean ± SD of three independent experiments. * p < 0.05 and ** p < 0.01 vs . control.

Article Snippet: The human AML stem cell line KG1a (#CCL-246.1) and Caucasian promyelocytic leukemia cell line HL60 (#CCL-240) were provided by the American Type Culture Collection (Manassas, VA, USA).

Techniques: Activity Assay, Membrane, Incubation, Staining, Control

Journal: PLoS ONE

Article Title: Blockade of p38 MAPK overcomes AML stem cell line KG1a resistance to 5-Fluorouridine and the impact on miRNA profiling

doi: 10.1371/journal.pone.0267855

Figure Lengend Snippet: Profiling of the top differentially dysregulated drug resistance-associated miRNA in AML stem cell line KG1a exposed to SB202190 p38 MAPK inhibitor and to 1–10 μM FUrd.

Article Snippet: The human AML stem cell line KG1a (#CCL-246.1) and Caucasian promyelocytic leukemia cell line HL60 (#CCL-240) were provided by the American Type Culture Collection (Manassas, VA, USA).

Techniques: Control

Journal: PLoS ONE

Article Title: Blockade of p38 MAPK overcomes AML stem cell line KG1a resistance to 5-Fluorouridine and the impact on miRNA profiling

doi: 10.1371/journal.pone.0267855

Figure Lengend Snippet: The KG1a cells were non-treated or treated with 10 μM SB202190 for 2 h incubation prior to cell treatment with 10 μM FUrd. After 48 h incubation, the total RNA was extracted for the miRNA RT-qPCR assay. The bar graphs show the relative expression level of miR-328-3p, miR-210-5p and miR-26b-5p mRNA determined by RT-qPCR analysis in non-treated and treated KG1a cells calculated as a ratio of the expression to the endogenous control miRNA. The results are presented as mean ± SD of three independent experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs . control.

Article Snippet: The human AML stem cell line KG1a (#CCL-246.1) and Caucasian promyelocytic leukemia cell line HL60 (#CCL-240) were provided by the American Type Culture Collection (Manassas, VA, USA).

Techniques: Incubation, miRNA RT, Expressing, Quantitative RT-PCR, Control