cellmask actin deep red actin tracking stain #a57245  (Thermo Fisher)

Journal: iScience

Article Title: Human monocyte-derived macrophages shift subcellular metalloprotease activity depending on their activation state

doi: 10.1016/j.isci.2024.111171

Figure Lengend Snippet:

Article Snippet: CellMask™ Deep Red Actin Tracking Stain , Invitrogen , Cat # A57245.

Techniques: Plasmid Preparation, Recombinant, Knock-Out, Blocking Assay, Staining, Antibody Labeling, Reverse Transcription, Lysis, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Fractionation, Cell Culture, Software, Flow Cytometry, Fluorescence, Spectrophotometry

Journal: bioRxiv

Article Title: A multicellular actin star network underpins epithelial organization and connectivity

doi: 10.1101/2024.07.26.605277

Figure Lengend Snippet: (A) Confocal analysis of myosin-IIA-KI-GFP (magenta) localization in AcSs (green). Scale bar, 5 μm. (B) Statistical analyses of the signal intensity level of myosin-IIA-GFP in nodes and cables of AcSs. Normalized myosin-IIA-GFP signal intensity in AcS nodes = 0.666±0.01 (mean±S.E.M), in AcS cables = 0.034±0.001. N = 3 experiments, n = 154 cells. Mann-Whitney test, ****p<0.0001. (C) N-SIM analysis of P-MLC2 (magenta) localization in AcSs (green). Scale bar, 5 μm. (D) Statistical analyses of the signal intensity level of P-MLC2 in nodes and cables of AcSs. P-MLC2 signal intensity in AcS nodes = 0.743±0.038 (mean±S.E.M), in cables = 0.417±0.026. N = 3 experiments, n = 98 cells. Unpaired t-test, ****p<0.0001. (E) Confocal analysis of the apico-basal distribution of myosin-IIA-GFP (green) or P-MLC2 (magenta) in organoid-derived monolayers. Scale bar, 10 μm. (F) Statistical analyses of the signal intensity level of myosin-IIA-GFP and P-MLC2 in the apical and basal domain of differentiated cells. Mean apical myosin-IIA-GFP signal intensity in differentiated compartments = 725±48 (mean±S.E.M), basal myosin-IIA-GFP = 1159±72, apical P-MLC2 = 1314±63, basal P-MLC2 = 2078±135. N = 3 experiments, n (cells in differentiated compartments) = 23 cells. Two-way ANOVA, ****p<0.0001. (G) Confocal analysis of myosin-IIA-GFP (green) and P-MLC2 (magenta) distribution in an organoid-derived monolayer. Nuclei (DNA) are stained with Hoechst33342 (blue). Crypt-like domains (c) are delimited with a dotted blue line. Scale bar, 20 μm. (H) Statistical analyses of the relative signal intensity level of myosin-IIA-GFP and P-MLC2 in differentiated and crypt-like compartments. Mean myosin-IIA-GFP signal intensity in differentiated compartments = 2049±128 (mean±S.E.M), in crypt-like compartments = 1572±163, mean P-MLC2 signal intensity in differentiated compartments = 3644±224, in crypt-like compartments = 1536±176. N = 3 experiments, n (cells in differentiated compartments) = 23 cells, n (cells in crypt-like compartments) = 19 cells. Two-way ANOVA, *p = 0.012, ****p<0.0001. (I) Confocal analysis and z-projection of actin distribution in the basal domain of control or blebbistatin-treated organoid-derived monolayers. Scale bar, left panel 20 μm, right panel 10 μm. (J) Time-lapse images of CellMask actin (green) in tdTomato (magenta) organoid-derived monolayer after 1h blebbistatin treatment and then wash-out (t = 0min). Crypt-like domains (c) are delimited with a dotted blue line. Scale bar, 10 μm. (K) Statistical analysis of the mean time of AcS re-formation after blebbistatin treatment and wash-out. Mean time (min) = 51.41±1.67 (mean±S.E.M). n = 101 cells.

Article Snippet: CellMask Actin Deep Red actin tracking stain (#A57245) was from Invitrogen (Thermo Fisher Scientific).

Techniques: MANN-WHITNEY, Derivative Assay, Staining, Control