Journal:

Article Title: Reinitiation of DNA Synthesis and Cell Division in Senescent Human Fibroblasts by Microinjection of Anti-p53 Antibodies

doi:

Figure Lengend Snippet: Morphological reversion and reinitiation of DNA synthesis in senescent human fibroblasts induced by anti-p53 antibody PAb1801. Senescent HCA2 cells (corresponding to PDL65 plus 19 days in Table ​Table1)1) were microinjected with control mouse IgG (B, E, and F), PAb1801 (C, G, and H), or plasmid SVori− (D) and analyzed 72 h later (all cells in the fields shown were injected). (B to D) Effects on morphology determined by phase-contrast microscopy compared to untreated young cultures (A). (E to H) Effects on DNA synthesis as revealed by BrdU labelling. Injected cells are identified by immunofluorescence of coinjected rat IgG with a rhodamine label (red) (E and G); nuclear BrdU incorporation (examples are indicated by arrows) is shown by immunofluorescence with FITC (green) (F and H). (Note that the weak cytoplasmic FITC fluorescence visible in panels F and H is due to detection of the injected mouse IgGs by the rabbit anti-mouse–FITC used to detect the BrdU.) Magnification, ×160.

Article Snippet: After being washed, cells were incubated as appropriate in secondary antibody (rhodamine-conjugated goat anti-rabbit IgG or anti-mouse IgG [Southern Biotechnology]) at 1:100 in PBS-bovine serum albumin for 1 h. Fluorescein isothiocyanate (FITC)-conjugated goat anti-rat IgG (Southern Biotechnology) was also added simultaneously to detect microinjected rat IgG and hence permit independent identification of microinjected cells.

Techniques: DNA Synthesis, Plasmid Preparation, Injection, Microscopy, Immunofluorescence, BrdU Incorporation Assay, Fluorescence

Journal:

Article Title: Reinitiation of DNA Synthesis and Cell Division in Senescent Human Fibroblasts by Microinjection of Anti-p53 Antibodies

doi:

Figure Lengend Snippet: Inhibition of p21sdi1/WAF1 expression in senescent fibroblasts by PAb1801 but not SV40. Senescent HCA2 cells (as for Fig. ​Fig.1)1) were microinjected with control IgG (A and B), PAb1801 (C and D), plasmid SVori− plus control IgG (E and F), or SVori− plus PAb1801 (G and H) and analyzed 72 h later by double immunofluorescence. Injected cells are shown by immunofluorescence detection of coinjected rat IgG with an FITC label (green) (A, C, E, and G). p21 content is shown by immunofluorescence with a rhodamine label (red) (B, D, F, and H). Examples of nuclei lacking detectable p21 after injection with PAb1801 are indicated with open arrows in panel D; a p21-positive nucleus in a neighboring uninjected cell is also visible (solid arrow). Note that PAb1801 (D) but not SVori− (F) causes loss of detectable nuclear p21 and that the persistent elevation in SVori−-injected cells is also abolished by coinjection of PAb1801 (H) (compare injected cells [open arrows] with uninjected cell [closed arrow]). Magnification, ×160.

Article Snippet: After being washed, cells were incubated as appropriate in secondary antibody (rhodamine-conjugated goat anti-rabbit IgG or anti-mouse IgG [Southern Biotechnology]) at 1:100 in PBS-bovine serum albumin for 1 h. Fluorescein isothiocyanate (FITC)-conjugated goat anti-rat IgG (Southern Biotechnology) was also added simultaneously to detect microinjected rat IgG and hence permit independent identification of microinjected cells.

Techniques: Inhibition, Expressing, Plasmid Preparation, Immunofluorescence, Injection

Journal: Bioactive materials

Article Title: Double hits with bioactive nanozyme based on cobalt-doped nanoglass for acute and diabetic wound therapies through anti-inflammatory and pro-angiogenic functions.

doi: 10.1016/j.bioactmat.2023.08.014

Figure Lengend Snippet: Fig. 2. CoNZ effects on anti-inflammation and pro-angiogenesis in full-thickness skin wound. (A) Illustration depicting the in vivo full-thickness skin wound model and experimental assays. (B) In vivo visualization of administered nanoparticles (CoNZ) by FITC-conjugation; at week 1, FITC-CoNZ nanoparticles were observed distributed within the wound area. (C) ROS signals (in red) in tissue samples at day 2 measured by dihydroethidium dye staining, along with semi-quantitative data using ImageJ; the intense ROS signal within the wound area was significantly reduced in the CoNZ group. (D) Immunohistochemical staining of tissue samples for iNOS (pro-inflammatory signal) and IL-10 (anti-inflammatory signal) at weeks 1 and 2, along with semi-quantitative data using ImageJ; iNOS was down-regulated while IL-10 was up-regulated in the CoNZ group. (E) Immunohistochemical staining of tissue samples for CD31 and α-SMA (neovascularization) at week 1, along with semi-quantitative data using ImageJ; both signals were expressed at significantly higher levels in the CoNZ group. (F) Western blot analysis of tissue samples for CD31, showing significant differences between groups (defect control < BGn « CoNZ ~ normal tissue); the dotted line (intensity level ‘1’) represents the normal group. Statistical significance was calculated between groups using one-way ANOVA with *p < 0.05, **p < 0.01, and ***p < 0.001 denoting significance compared to the defect group; and +p < 0.05, ++p < 0.01, and +++p < 0.001 indicating significance compared to the BGn group. All data are presented as mean ± one standard deviation and the sample size was n = 4.

Article Snippet: After washing the tissue sections three times for 5 min each, secondary antibodies conjugated with Rhodamine and FITC (1:200, Jackson Immuno Research, INC) or FITC (Rat, NB7114, Novus) were applied for 1 h at room temperature.

Techniques: In Vivo, Conjugation Assay, Staining, Immunohistochemical staining, Western Blot, Control, Standard Deviation