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Figure 2. The isoform-specific effects of decreased non-muscle actin expression on lamina composition in A549 cell line. (a,b) The effects of shRNA treatment targeting non-muscle actins (4 days) on lamins A/C in A549 cells, immunofluorescence microscopy. Lamins A/C—green, γ-actin—red, DNA— blue; (c) Western blot analysis of lamins A/C in A549 cell line (shRNA to β- or γ-actin, 4 days); (d,e) Lamin <t>B2</t> immunofluorescence depending on the suppression of β- or γ-actin in A549 cells (4 days). Lamin B2—green, γ -actin—red, DNA—blue; (f) Western blot analysis of lamin B2 in A549 cells (shRNA to β- or γ-actin, 4 days). Mann–Whitney U test was used for the statistical analysis for all the depicted comparisons. Asterisks indicate p-values <0.01 (**) for all panels. Scale bar 10 µm for all immunofluorescent microscopy panels.
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Figure 2. The isoform-specific effects of decreased non-muscle actin expression on lamina composition in A549 cell line. (a,b) The effects of shRNA treatment targeting non-muscle actins (4 days) on lamins A/C in A549 cells, immunofluorescence microscopy. Lamins A/C—green, γ-actin—red, DNA— blue; (c) Western blot analysis of lamins A/C in A549 cell line (shRNA to β- or γ-actin, 4 days); (d,e) Lamin B2 immunofluorescence depending on the suppression of β- or γ-actin in A549 cells (4 days). Lamin B2—green, γ -actin—red, DNA—blue; (f) Western blot analysis of lamin B2 in A549 cells (shRNA to β- or γ-actin, 4 days). Mann–Whitney U test was used for the statistical analysis for all the depicted comparisons. Asterisks indicate p-values <0.01 (**) for all panels. Scale bar 10 µm for all immunofluorescent microscopy panels.

Journal: International journal of molecular sciences

Article Title: Divergent Contribution of Cytoplasmic Actins to Nuclear Structure of Lung Cancer Cells.

doi: 10.3390/ijms252413607

Figure Lengend Snippet: Figure 2. The isoform-specific effects of decreased non-muscle actin expression on lamina composition in A549 cell line. (a,b) The effects of shRNA treatment targeting non-muscle actins (4 days) on lamins A/C in A549 cells, immunofluorescence microscopy. Lamins A/C—green, γ-actin—red, DNA— blue; (c) Western blot analysis of lamins A/C in A549 cell line (shRNA to β- or γ-actin, 4 days); (d,e) Lamin B2 immunofluorescence depending on the suppression of β- or γ-actin in A549 cells (4 days). Lamin B2—green, γ -actin—red, DNA—blue; (f) Western blot analysis of lamin B2 in A549 cells (shRNA to β- or γ-actin, 4 days). Mann–Whitney U test was used for the statistical analysis for all the depicted comparisons. Asterisks indicate p-values <0.01 (**) for all panels. Scale bar 10 µm for all immunofluorescent microscopy panels.

Article Snippet: The following primary antibodies were applied: - mouse monoclonal antibodies to pan-actin (clone C4, Cell Signaling Technology Inc., Danvers, MA, USA), β-actin (IgG1, MCA5775GA, AbD Serotec, Kidlington, UK), γ-actin (IgG2b, MCA5776GA, AbD Serotec), histone H3 (di methyl K9) (IgG2a, ab1220, Abcam, Cambridge, UK); - rabbit monoclonal antibodies to histone H1.4 (D4J5Q, Cell Signaling Technology Inc.), histone H2A (D6O3A, Cell Signaling Technology Inc.), histone H3 (4499, Cell Signaling Technology Inc.), lamin B2 (D8P3U, Cell Signaling Technology Inc.), lamin A/C (4C11, Cell Signaling Technology Inc.); Int.

Techniques: Expressing, shRNA, Immunofluorescence, Microscopy, Western Blot, MANN-WHITNEY

Figure 3. Laser scanning microscopy of lamin B2 immunofluorescence in A549 cells after shRNA targeting non-muscle actins treatment. (a) A comparative analysis of a control culture with shRNA derivatives (4 days). Lamin B2—green, β-actin—red, DNA—blue. Scale bar—10 µm; (b) Lamin B2 immunofluorescence in A549 treated with shRNA to β-actin (4 days). Lamin B2—green, DNA—blue. Scale bar—10 µm; (c) The mean IF intensity of lamin B2 in the nuclear envelope zone of A549 cells. Mann–Whitney U test was used for the statistical analysis for the depicted comparisons. Asterisks indicate p-values < 0.01 (**).

Journal: International journal of molecular sciences

Article Title: Divergent Contribution of Cytoplasmic Actins to Nuclear Structure of Lung Cancer Cells.

doi: 10.3390/ijms252413607

Figure Lengend Snippet: Figure 3. Laser scanning microscopy of lamin B2 immunofluorescence in A549 cells after shRNA targeting non-muscle actins treatment. (a) A comparative analysis of a control culture with shRNA derivatives (4 days). Lamin B2—green, β-actin—red, DNA—blue. Scale bar—10 µm; (b) Lamin B2 immunofluorescence in A549 treated with shRNA to β-actin (4 days). Lamin B2—green, DNA—blue. Scale bar—10 µm; (c) The mean IF intensity of lamin B2 in the nuclear envelope zone of A549 cells. Mann–Whitney U test was used for the statistical analysis for the depicted comparisons. Asterisks indicate p-values < 0.01 (**).

Article Snippet: The following primary antibodies were applied: - mouse monoclonal antibodies to pan-actin (clone C4, Cell Signaling Technology Inc., Danvers, MA, USA), β-actin (IgG1, MCA5775GA, AbD Serotec, Kidlington, UK), γ-actin (IgG2b, MCA5776GA, AbD Serotec), histone H3 (di methyl K9) (IgG2a, ab1220, Abcam, Cambridge, UK); - rabbit monoclonal antibodies to histone H1.4 (D4J5Q, Cell Signaling Technology Inc.), histone H2A (D6O3A, Cell Signaling Technology Inc.), histone H3 (4499, Cell Signaling Technology Inc.), lamin B2 (D8P3U, Cell Signaling Technology Inc.), lamin A/C (4C11, Cell Signaling Technology Inc.); Int.

Techniques: Laser-Scanning Microscopy, Immunofluorescence, shRNA, Control, MANN-WHITNEY