Journal: Oncotarget

Article Title: Identification of a novel functional JAK1 S646P mutation in acute lymphoblastic leukemia

doi: 10.18632/oncotarget.16670

Figure Lengend Snippet: ( A ) The proliferation of transduced BaF3 cells after withdrawing IL-3 for 3 days was assayed by CCK-8. Data are expressed as mean ± s.d of at least 3 independent experiments. ** P < 0.01. ( B ) The viability of transduced BaF3 cells after withdrawing IL-3 for 48 h was determined by trypan blue exclusion method at the indicated time points. ( C ) The survival curves of transduced BaF3 cells cultured in the absence of IL-3 for 6 days were determined by CCK-8. ( D ) Cell cycle histograms of transduced BaF3 cells were assayed by flow cytometry. The proportion of G1 or S/G2 phase is expressed as percentage of total cells.

Article Snippet: BaF3 murine pro-B cells was obtained from the China Infrastructure of Cell Line Resources, cultured in RPMI 1640 medium (Hyclone, Logan, Utah, USA) with 10% fetal bovine serum (FBS) (Gibco, California, USA) and 10 ng/ml mIL-3 (Peprotech, Rocky Hill, USA).

Techniques: CCK-8 Assay, Cell Culture, Flow Cytometry

Journal: Oncotarget

Article Title: Identification of a novel functional JAK1 S646P mutation in acute lymphoblastic leukemia

doi: 10.18632/oncotarget.16670

Figure Lengend Snippet: 2 × 10 6 BaF3 cells transduced with vector, WT JAK1 , or S646P mutant were intravenously inoculated into nude mice. ( A ) WBC counts were monitored at the indicated time points. ( B ) GFP-positive cells in peripheral blood were monitored by flow cytometry. ( C ) At the time of sacrifice, the morphological changes of livers and spleens were photographed. ( D , E ) The weights of livers (D) and spleens (E) were recorded. Data are expressed as the mean± s.d. * P < 0.05. ( F ) The bone marrow smears were stained with Wright and photographed (magnification: ×1000). Representative abnormal cells were marked with arrows. ( G ) The percentage of GFP-positive cells in bone marrow at the time of sacrificed was detected by flow cytometry. Data are expressed as the mean ± s.d. * P < 0.05.

Article Snippet: BaF3 murine pro-B cells was obtained from the China Infrastructure of Cell Line Resources, cultured in RPMI 1640 medium (Hyclone, Logan, Utah, USA) with 10% fetal bovine serum (FBS) (Gibco, California, USA) and 10 ng/ml mIL-3 (Peprotech, Rocky Hill, USA).

Techniques: Transduction, Plasmid Preparation, Mutagenesis, Flow Cytometry, Staining

Journal: Oncotarget

Article Title: Identification of a novel functional JAK1 S646P mutation in acute lymphoblastic leukemia

doi: 10.18632/oncotarget.16670

Figure Lengend Snippet: ( A ) Transiently transfected HEK293V cells were treated with increasing concentrations of ruxolitinib for 24 h. Cell lysates were immunoblotted with specific antibodies shown. GAPDH was used as loading control. ( B ) Transduced BaF3 cells were treated with increasing concentrations of ruxolitinib for 48 h. Cell proliferation was determined by CCK-8; K562 cells were used as negative control. Results represent the mean ± s.d. of 3 independent experiments.

Article Snippet: BaF3 murine pro-B cells was obtained from the China Infrastructure of Cell Line Resources, cultured in RPMI 1640 medium (Hyclone, Logan, Utah, USA) with 10% fetal bovine serum (FBS) (Gibco, California, USA) and 10 ng/ml mIL-3 (Peprotech, Rocky Hill, USA).

Techniques: Transfection, CCK-8 Assay, Negative Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Mitochondrial Targeted Antioxidant SKQ1 Ameliorates Acute Kidney Injury by Inhibiting Ferroptosis

doi: 10.1155/2022/2223957

Figure Lengend Snippet: SKQ1 suppresses CDDP-induced mitochondrial oxidative stress and ferroptosis in HK2 cells. (a) Viability of HK2 cells after treatment with various concentrations of SKQ1. (b) The relative MitoSOX levels of HK2 cells treated with CDDP (10 μ g/mL) with or without 20 nM SKQ1 were then analyzed by flow cytometry. Representative fluorescence images of (c) MitoSOX and (d) TMRM in the four different groups of HK2 cells (red, MitoSOX or TMRM; blue, Hoechst). (e) Mitochondrial lipid peroxidation was evaluated by staining with MitoPeDPP and MFI of oxidized MitoPeDPP and analyzed using flow cytometry. (f) Representative fluorescence images of lipid peroxidation after cell staining with BODIPY dye (red, lipids; green, oxidized lipids; blue, Hoechst). (g) LDH release from HK2 cells for all groups, and (h) relative levels of MDA and the (i) relative GSH/GSSG ratio in HK2 cells treated with CDDP and cotreated with vehicle or SKQ1. (j) We applied a CCK-8 assay to ascertain the viability of HK2 cells treated with SKQ1, 10 μ M liproxstatin-1, or 20 mM Z-VAD-FMK, respectively, or with a combination of all three, followed by treatment with CDDP. (k) Immunoblots of GPX4 and cleaved caspase-3 in HK2 cells after treatment with CDDP and cotreatment with vehicle or SKQ1. Three independent assays were conducted. ∗∗∗∗ P < 0.0001, ∗∗∗ P < 0.001, ∗∗ P < 0.01, and ∗ P < 0.05 (by one-way or two-way ANOVA).

Article Snippet: Sources and authentication of human proximal tubule epithelial cells (HK2) were from the American Type Culture Collection.

Techniques: Flow Cytometry, Fluorescence, Staining, CCK-8 Assay, Western Blot

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: MicroRNA-613 is downregulated in HCMV-positive glioblastoma and inhibits tumour progression by targeting arginase-2.

doi: 10.1177/1010428317712512

Figure Lengend Snippet: Figure 4. MiR-613 negatively regulates ARG2 expression in GBM cells. (a) ARG2 protein level was detected by western blot in GBM cells after transfection. GAPDH was used as an internal control. (b) The 3′-UTR sequences of wild-type/mutant ARG2 were cloned into the pMIR-report luciferase vector. The predicted binding sites between miR-613 and the 3′-UTR sequence of ARG2 is shown. (c and d) GBM cells were co-transfected with the luciferase reporter plasmid carrying the 3′-UTR sequences of wild-type/ mutant ARG2 and control/miR-613 mimics. Luciferase activity was analysed 48 h after co-transfection using dual-luciferase assays (*p < 0.05, n = 5).

Article Snippet: Human polyclonal antibody ARG2 (catalog number: 19324, final dilution 1:1000, Source: Rabbit; Cell Signaling Technology, Boston, MA, USA) was used as the primary antibody.

Techniques: Expressing, Western Blot, Transfection, Control, Mutagenesis, Clone Assay, Luciferase, Plasmid Preparation, Binding Assay, Sequencing, Activity Assay, Cotransfection

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: MicroRNA-613 is downregulated in HCMV-positive glioblastoma and inhibits tumour progression by targeting arginase-2.

doi: 10.1177/1010428317712512

Figure Lengend Snippet: Figure 5. Re-expression of ARG2 reverses the inhibitory properties of miR-613. U87 and U251 cells were co-transfected with control/miR-613 mimics and the empty vector/ARG2 recombinant plasmid for 48 h. (a) The expression of ARG2 protein was analysed by western blot. GAPDH was used as an internal control. (b) Cell growth curves for GBM cells were measured by the CCK-8 assay. (c) Cell apoptosis was analysed by flow cytometry. (d) Clone-formation assay. The number of colonies was quantified. (e) Transwell assays. The number of invasive cells was quantified. (f) Wound-healing assays. The percentage of the distance migrated was quantified (*p < 0.05, n = 5).

Article Snippet: Human polyclonal antibody ARG2 (catalog number: 19324, final dilution 1:1000, Source: Rabbit; Cell Signaling Technology, Boston, MA, USA) was used as the primary antibody.

Techniques: Expressing, Transfection, Control, Plasmid Preparation, Recombinant, Western Blot, CCK-8 Assay, Flow Cytometry, Tube Formation Assay