Journal: Scientific reports

Article Title: Administration of an antibody against apoptosis inhibitor of macrophage prevents aortic aneurysm progression in mice.

doi: 10.1038/s41598-024-66791-7

Figure Lengend Snippet: Figure 5. AIM distribution. (A,B) Representative images of immunofluorescence staining for AIM in the IgG and AIM groups. AIM (red) and cell nuclei (blue). Scale bars = 50 μm. (C) The ratio of the AIM-positive area to the DAPI-positive area, expressed as a percentage. Data are means ± SEM. **p < 0.01 assessed by the Mann– Whitney U test. The “#” symbols enclosed by dashed lines indicate the vessel lumens.

Article Snippet: In all slides, cell nuclei were stained with DAPI Fluoromount-G (Southern Biotech, Birmingham, AL, USA).

Techniques: Immunofluorescence, Staining, MANN-WHITNEY

Journal: Scientific reports

Article Title: Administration of an antibody against apoptosis inhibitor of macrophage prevents aortic aneurysm progression in mice.

doi: 10.1038/s41598-024-66791-7

Figure Lengend Snippet: Figure 6. Macrophage polarization. (A) iNOS (red) and CD68 (green); (B) CD206 (red), CD68 (green), and cell nuclei (blue). Scale bars = 100 μm. (C) The percentage of CD68 + cells to DAPI + cells. (D,E) The percentage of the CD68 + population consisting of iNOS + /CD206 + cells. (F) The percent ratio of CD206 + cells to iNOS + cells in the CD68 + population. Data are means ± SEM. *p < 0.05 and **p < 0.01 assessed by the Mann– Whitney U test. iNOS + /CD68 + cells indicate M1 macrophages, while CD206 + /CD68 + cells indicate M2 macrophages. The “#” symbols enclosed by dashed lines indicate the vessel lumens.

Article Snippet: In all slides, cell nuclei were stained with DAPI Fluoromount-G (Southern Biotech, Birmingham, AL, USA).

Techniques: MANN-WHITNEY

Journal: Life

Article Title: Anticancer Potential of Pyridoxine-Based Doxorubicin Derivatives: An In Vitro Study

doi: 10.3390/life14030282

Figure Lengend Snippet: The distribution of MCF-7 cells into apoptosis phases after DOX-2 treatment for 72 h according to Annexin V-FITC/DAPI staining. Treatment with Triton X-100—24 h. The figures present the average data derived from two independent experiments conducted in duplicate.

Article Snippet: Then, cells were washed, placed in a binding buffer, stained with Annexin V-FITC/DAPI, and analyzed using the flow cytofluorimeter BD FACSCalibur™ (BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Staining, Derivative Assay

Journal: PLoS ONE

Article Title: Depletion of Nuclear Poly(A) Binding Protein PABPN1 Produces a Compensatory Response by Cytoplasmic PABP4 and PABP5 in Cultured Human Cells

doi: 10.1371/journal.pone.0053036

Figure Lengend Snippet: HeLa cells were grown on 35 mm plates and transiently transfected with PABPN1 Si-RNA or PABPN-UTR RNAi (A) Sixty six hours after transfection, cytoplasmic and nuclear extracts were prepared from UTR, and Si-PABPN1 cells and probed with PABP1, 3, 4, 5, and PABPN1 antibodies. GAPDH and SKIP (loading control) were used to determine cross contamination in each fractions. (B) Nuclear fractions from non transfected (NT), PABPN1-Si and UTR-Si transfected cells were used for western blotting with PABP4, PABPN1, and SKIP (loading control) antibodies as previously described. (C) Nuclear PABP4 abundance was calculated by scanning band intensities and expressed as an arbitrary unit. Data are mean ± SE, P<0.05, n = 3. (D) Following 66 h of transfection with Si RNAs HeLa cells grown on cover slips were fixed with Para-formaldehyde and processed for immunostaining with PABP1, 4, 5 specific antibodies and counterstained with Texas-red-conjugated secondary antibody. (E) The PABP4 stained slide was further stained with DNA staining dye, DAPI, to show the nuclear localization of PABP4 in PABPN1-Si transfected cells. Processed specimens were then examined with a confocal microscope (F) The nuclear fractions of NT, PABPN1-UTR and PABPN1-Si transfected cells after 66 h of treatment were immunoprecipitated with PABP4 antibody; the RNA bound to PABP4 was extracted, and analyzed by RT-PCR with β-actin pre-mRNA specific primers as described in legends to . The band intensities are shown in arbitrary unit. Immunoprecipitation with PABPN1 antibody was used as a positive control. The control lane indicate PCR product of immunoprecipitated RNA from PABPN1-Si cells without reverse transcription. The lanes at the bottom panel show β-actin pre-mRNA immunoprecipitated with PABPN1 antibody from PABPN1-UTR and PABPN1-Si transfected cells as controls. (G) Immunoprecipitation from the nuclear extract was carried out with PABP4 antibody as described in . The agarose bead eluted (IP-PABP4) and cell equivalent amount of the input nuclear extract were used for western blotting (WB) with CPSF and PABP4 antibody.

Article Snippet: After immunostaining with PABP4 antibody, the nuclei of the cells were further stained with DAPI (Santa Cruz Biotechnology, CA, USA) (0.5 μg/μl) for 10 min, washed twice with PBS and mounted onto slides.

Techniques: Transfection, Control, Western Blot, Immunostaining, Staining, Microscopy, Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction, Positive Control, Reverse Transcription