American Society of Hematology

Fig. 4.

$Fig. 4. Effect of scinderin expression and its activation by ionophore A-23187 on filamentous actin content in MEG-01 cells. / (A) Fluorescence microscope images of cells stained with scinderin (Sc) antibodies and rhodamine phalloidin (a probe for F-actin) after transfection either with vector pcDNA3 (2 left panels) or with Sc cDNA carrying vector pcDNA3-Sc (2 right panels). (B) Image analysis data of the fluorescent cells described in panel A. Twelve preparations of pcDNA3-transfected cells were analyzed, and 18 preparations from 3 different clones (6 from each clone) were measured for pcDNA3-Sc–transfected cells (*P < .001). (C) Western blot analysis carried out with an actin antibody on supernatants (S) and sediments of Triton X-100 extracts prepared from wild-type MEG-01 cells (W.T.) and from cells transfected with either pcDNA3 or pcDNA3-Sc. A decrease in the Triton X-100–insoluble (I) fraction (filamentous actin) was observed in pcDNA3-Sc–transfected cells. (D) Two-minute treatment with ionophore A23187 further decreased levels of F-actin in pcDNA3-Sc–transfected cells. F-actin was measured as described in panel B. Bars represent mean ± SEM from 4 to 5 different preparations, each containing 20 to 35 cells (**P < .05). (E) Fluorescence microscope images of a sample of cells described in panel D after rhodamine phalloidin staining.$

Effect of scinderin expression and its activation by ionophore A-23187 on filamentous actin content in MEG-01 cells.

(A) Fluorescence microscope images of cells stained with scinderin (Sc) antibodies and rhodamine phalloidin (a probe for F-actin) after transfection either with vector pcDNA3 (2 left panels) or with Sc cDNA carrying vector pcDNA3-Sc (2 right panels). (B) Image analysis data of the fluorescent cells described in panel A. Twelve preparations of pcDNA3-transfected cells were analyzed, and 18 preparations from 3 different clones (6 from each clone) were measured for pcDNA3-Sc–transfected cells (*P < .001). (C) Western blot analysis carried out with an actin antibody on supernatants (S) and sediments of Triton X-100 extracts prepared from wild-type MEG-01 cells (W.T.) and from cells transfected with either pcDNA3 or pcDNA3-Sc. A decrease in the Triton X-100–insoluble (I) fraction (filamentous actin) was observed in pcDNA3-Sc–transfected cells. (D) Two-minute treatment with ionophore A23187 further decreased levels of F-actin in pcDNA3-Sc–transfected cells. F-actin was measured as described in panel B. Bars represent mean ± SEM from 4 to 5 different preparations, each containing 20 to 35 cells (**P < .05). (E) Fluorescence microscope images of a sample of cells described in panel D after rhodamine phalloidin staining.

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