cellrox green flow cytometry kit  (Thermo Fisher)

Journal: Cells

Article Title: Evaluation of Anti-Inflammatory Activity of the New Cardiotonic Steroid γ-Benzylidene Digoxin 8 (BD-8) in Mice

doi: 10.3390/cells13181568

Figure Lengend Snippet: Effect of BD-8 on phagocytosis. Murine peritoneal macrophages were treated with 10 µM BD-8. After 24 h of incubation, the cells were stimulated with 1:10 red ZYM (representative fluorescence dot plots ( A ) and incubated again for 40 min and the phagocytosis was evaluated. Flow cytometry data were analyzed using FlowJo ® X software version 10.6.2. ( B ) Results were expressed as mean ± SEM and analyzed using GraphPad Prism ® software version 10.0.0 using one-way analysis of variance followed by Tukey’s test for multiple comparisons. ****—( p < 0.0001); *—( p = 0.05). CTR: Control; RedZym: Red ZYM.

Article Snippet: Similarly, ROS production was assessed accordingly CellROX™ Green Flow Cytometry Assay Kit (C10492 Invitrogen™).

Techniques: Incubation, Fluorescence, Flow Cytometry, Software, Control

Journal: Cells

Article Title: Evaluation of Anti-Inflammatory Activity of the New Cardiotonic Steroid γ-Benzylidene Digoxin 8 (BD-8) in Mice

doi: 10.3390/cells13181568

Figure Lengend Snippet: Effects of BD-8 on intracellular molecules associated with inflammatory processes and histograms. Peritoneal macrophages were stimulated with 0.2 mg/mL ZYM and treated with 10 µM BD-8. The cells were evaluated by flow cytometry and marked with specific antibodies for intracellular molecules. ( A ) p-ERK+; ( B ) p-p38+; ( C ) p-NF- κB+; ( D ) p-Akt+; ( E ) p-mTOR+; ( F ) ROS+; ( G ) iNOS+; ( H ) COX-2+; ( I ) Src+. The numerical data are presented as mean ± SEM with an average of n = 5 and were analyzed using one-way analysis of variance (ANOVA), followed by Tukey’s post-test. # p < 0.05, significant in relation to the control group; * p < 0.05, significant in relation to the ZYM group. CTR: Control; ZYM: Zymosan; ns: not significant.

Article Snippet: Similarly, ROS production was assessed accordingly CellROX™ Green Flow Cytometry Assay Kit (C10492 Invitrogen™).

Techniques: Flow Cytometry, Control

Journal: Molecular Medicine

Article Title: Modulation of anti-cardiac fibrosis immune responses by changing M2 macrophages into M1 macrophages

doi: 10.1186/s10020-024-00858-z

Figure Lengend Snippet: NaW increases lysosomal pH-mediated Ca 2+ release and thus enhances glucose uptake. A – G IL-4-conditioned bone marrow-derived macrophages (BMDMs) were treated with or without NaW (2 mM) for 12 h. (A) Scl2a1 expression was measured by real-time PCR. B The mean fluorescence intensity (MFI) level of 2NBDG was measured by flow cytometry. C LysoSensor Green-labeled acidic lysosomes were observed under a fluorescence microscope. Scale bar, 20 μm. D The MFI of the LysoSensor probe was measured by flow cytometry. E , F TMEM175 expression was measured by real-time PCR ( E ) and western blotting ( F ). G – I IL-4-conditioned BMDMs were transfected with TMEM175 siRNAs for 12 h, the MFI of the LysoSensor probe was detected by flow cytometry ( G ), and Nos2 expression was determined by real-time PCR ( H ) and western blotting ( I ). J After Tmem175 overexpression in macrophages, the MFI emitted by 2NBDG was measured by flow cytometry. K IL-4-conditioned BMDMs were treated with or without NaW (2 mM) for 12 h, and cytosolic calcium release was measured by flow cytometry. L , M IL-4-conditioned BMDMs were treated with NH 4 Cl for 5 min, the pH value of lysosomes was measured with a microplate reader ( L ), and the cytosolic calcium release rate was measured by flow cytometry ( M ). N The expression of Mcoln1 and Mcoln2 was analyzed by real-time PCR. O The same as presented in ( A ), except cells were pretreated with 10 nM CsA for 1 h. P , Q The same as presented in ( A ), except the expression and location of TFEB were analyzed by real-time PCR ( P ) and immunofluorescence ( Q ) (scale bar, 20 μm). R , S IL-4-conditioned BMDMs were transfected with Tfeb siRNA, Scl2a1 expression was measured by real-time PCR, and the MFI of 2NBDG was measured by flow cytometry. Unless otherwise specified, n = 3 biologically independent experiments. The data are presented as the mean ± SEM. P values were calculated using one-way ANOVA. *P < 0.05; **P < 0.01; ***P < 0.001

Article Snippet: ROS levels were measured using CellROX Green flow cytometry assay kits (C10444, Invitrogen).

Techniques: Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Fluorescence, Flow Cytometry, Labeling, Microscopy, Western Blot, Transfection, Over Expression, Immunofluorescence