prolong diamond with dapi counterstaining  (Fisher Scientific)

Journal: Cells

Article Title: Targeting of p21-Activated Kinase 4 Radiosensitizes Glioblastoma Cells via Impaired DNA Repair

doi: 10.3390/cells11142133

Figure Lengend Snippet: PAK isoforms differentially regulate glioblastoma cell radiosensitivity. ( A ) Representative Western blot images of the expression of the indicated PAK isoforms in LN229 and U343MG cell lines. β-actin served as loading control. ( B ) Representative immunofluorescence images of the nucleus (DAPI), the indicated PAK isoforms, and F-actin (Phalloidin) in LN229 cells. Scale bar: 5 µm. ( C ) Workflow of the colony formation assay for the quantification of clonogenic survival. ( D , E ) Normalized plating efficiency (basal clonogenic survival) of ( D ) LN229 and ( E ) U343MG cells upon knockdown of the different PAK isoforms. ( F , G ) Surviving fraction of ( F ) LN229 and ( G ) U343MG cells upon knockdown of the different PAK isoforms with and without 4 Gy X-ray irradiation. ( D – G ) Data show means +/– SDs (N = 3 independent experiments, one-way ANOVA, Dunnett’s post-hoc test, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Cells were mounted with ProLong Diamond Antifade Mountant with DAPI counterstain (Thermo Fisher Scientific).

Techniques: Western Blot, Expressing, Control, Immunofluorescence, Colony Assay, Knockdown, Irradiation

Journal: Cells

Article Title: Targeting of p21-Activated Kinase 4 Radiosensitizes Glioblastoma Cells via Impaired DNA Repair

doi: 10.3390/cells11142133

Figure Lengend Snippet: PAK4 depletion triggers an accumulation of DNA double-strand breaks. ( A , B ) Representative immunofluorescence images of the nucleus (DAPI), γH2AX, and 53BP1 in LN229 and U343MG cells upon treatment with ( A ) siPAK4#1 and ( B ) PAK4i EC50 without or in combination with 6 Gy X-ray irradiation. Scale bar: 5 µm. ( C , D ) Quantification of γH2AX/53BP1 foci in LN229 and U343MG cells upon ( C ) siRNA-mediated depletion of PAK4 and ( D ) PAK4i treatment in unirradiated and 6 Gy X-ray irradiated cells. ( E , F ) Proportion of cells with indicated numbers of γH2AX/53BP1 foci upon ( E ) depletion of PAK4 and ( F ) PAK4i treatment before and after 6 Gy X-ray irradiation. ( C – F ) Data show means +/− SDs (N = 3 independent experiments, ∑n = 150 cells per treatment condition), one-way ANOVA, Bonferroni’s post-hoc test, *** p < 0.001).

Article Snippet: Cells were mounted with ProLong Diamond Antifade Mountant with DAPI counterstain (Thermo Fisher Scientific).

Techniques: Immunofluorescence, Irradiation