human t-cell lymphoblastic leukemia cell line jurkat  (Procell Inc)

Journal: Pathology and Oncology Research

Article Title: Knockdown of GPSM1 Inhibits the Proliferation and Promotes the Apoptosis of B-Cell Acute Lymphoblastic Leukemia Cells by Suppressing the ADCY6-RAPGEF3-JNK Signaling Pathway

doi: 10.3389/pore.2021.643376

Figure Lengend Snippet: Downregulation of GPSM1 suppressed cell proliferation, induced cell cycle arrest and promoted apoptosis in BALL-1 and Reh cells. (A) Expression of GPSM1 mRNA in the human B lymphoblast cell line HMy2.CIR and several human leukemia cell lines (BALL-1, Jurkat and Reh). GAPDH was used as an internal reference for normalization. * p < 0.05 vs. the HMy2.CIR group. (B) GPSM1 protein expression in the human B lymphoblast cell lineHMy2.CIR and several human leukemia cell lines (BALL-1, Jurkat and Reh) was investigated by western blotting. (C) The grayscale value of the GPSM1 protein band was quantified by Tanon Image software, and GAPDH was used as an internal reference. * p < 0.05 vs. the HMy2.CIR group. (D) Relative mRNA expression in sh-Con- and sh-GPSM1-transfected BALL-1 and Reh cells. (E) GPSM1 protein expression in sh-Con- and sh-GPSM1-transfected BALL-1 and Reh cells. (F) The grayscale value of the GPSM1 protein band was quantified by Tanon Image software, and GAPDH was used as an internal reference. * p < 0.05, compared with the sh-Con group. (G) The CCK-8 assay was used to evaluate the cell proliferation ability of BALL-1 and Reh cells transfected with sh-GPSM1 and sh-Con on four consecutive days. * p < 0.05, ** p < 0.01, compared with the sh-Con group. (H,I) The cell cycle was examined by PI staining using flow cytometry. (J) Cell apoptosis rates were assessed by flow cytometry after staining cells with annexin V-PE/7-AAD. Nonapoptotic cells: annexin V-PE − /7-AAD − , early apoptotic cells: annexin V-PE + /7-AAD − , late apoptotic cells: annexin V-PE + /7-AAD + . PE, phycoerythrin; 7-AAD, 7-amino-actinomycin D. (K) The total apoptosis rate is the sum of the early apoptosis rate and the late apoptosis rate. * p < 0.05, compared with the sh-Con group. All data are expressed as the mean ± SD.

Article Snippet: A human T-cell lymphoblastic leukemia cell line (Jurkat) was obtained from Procell Life Science and Technology Co., Ltd. A human lymphoblastic leukemia cell line (Reh) was obtained from Shanghai Genechem Co., Ltd (Shanghai, China).

Techniques: Expressing, Western Blot, Software, Transfection, CCK-8 Assay, Staining, Flow Cytometry

Journal: BMC Microbiology

Article Title: IRAK-M alters the polarity of macrophages to facilitate the survival of Mycobacterium tuberculosis

doi: 10.1186/s12866-017-1095-2

Figure Lengend Snippet: IRAK-M over-expression resulted in higher bacterial load, while IRAK-M interference resulted in lower bacterial load in M. tb infected cells. Lentiviruses that over-expresses (IRAK-M-GFP-Lentivirus, OE) or interferes with (IRAK-M-RNAi-GFP-Lentivirus, KD) IRAK-M expression, were constructed. a Lentiviruses of negative control (NC) and OE were introduced into Jurkat cells. Cells were examined by light microscope and fluorescent microscope at 96 h post-transfection. More than 80% of Jurkat cells expressed GFP (200×, scale bar 50 μm). b Lentiviruses of NC and KD were introduced in U937 cells. Cells were examined by light microscope and fluorescent microscope at 96 h post-transfection. More than 90% of U937 cells expressed GFP (200×, scale bar 100 μm). c Expression of IRAK1-4 in Jurkat and U937 cells was evaluated by Western blot. d - g Bacterial load in Jurkat ( d , e ) and U937 cells ( f , g ) challenged with virulent M. tuberculosis H37Rv strain (MOI 10) were analyzed by acid-fast staining and colony forming units (CFU). Cells were infected with lentivirus for 96 h, challenged with H37Rv for 5 h and washed three times with PBS. d , f At 24 h post-infection, 1 × 10 6 cells were resuspended in 20 μl PBS and fixed in 4% para-formaldehyde for 15 min on the slides. Acid-fast staining (AF) was performed and arrows indicated AF-positive bacteria in the cells (1000×, scale bar 10 μm). e , g For determination of CFU, at 5 and 24 h post-infection, 1 × 10 5 cells were washed aseptically, homogenized and plated at 10-fold serial dilutions on Middlebrook 7H11 agar. CFU numbers per plate were counted to evaluate the bacterial load 3 to 4 weeks later. The bacterial load in cells of different groups was expressed as Log10 CFU ± SEM (**, p < 0.01; ***, p < 0.001)

Article Snippet: Human monocytic leukemia line U937 (ATCC® CRL-1593.2) and human T-cell acute lymphoblastic leukemia line Jurkat (ATCC® TIB-152TM) were cultured at 37 °C in CO 2 incubator (CCL-170B-8, ESCO, Singapore) in RPMI-1640 medium, supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin.

Techniques: Over Expression, Infection, Expressing, Construct, Negative Control, Light Microscopy, Microscopy, Transfection, Western Blot, Staining, Bacteria