Journal: Biology Open

Article Title: P2X 7 regulates ependymo-radial glial cell proliferation in adult Danio rerio following spinal cord injury

doi: 10.1242/bio.060270

Figure Lengend Snippet: P2X 7 receptors were expressed by GFAP + ependymo-radial glial cells. Spinal cord cross sections are shown (dorsal is up; asterisk indicates central canal; scale bar: 50 μm). Representative images of GFAP and P2X 7 labelling in naïve fish (A-C′), at 7 dpi (D-F′), and at 14 dpi (G-I′). Colocalization (arrows) was seen in radial processes and cell bodies (C′,F′,I′). GFAP + cells that did not colocalize with P2X 7 were also detected (open arrowheads) (C′,F′,I′). n =3/group. Created with BioRender.com.

Article Snippet: This included a P2X 7 control antigen (Alomone Labs, BLP-PR004, Q64663).

Techniques:

Journal: Biology Open

Article Title: P2X 7 regulates ependymo-radial glial cell proliferation in adult Danio rerio following spinal cord injury

doi: 10.1242/bio.060270

Figure Lengend Snippet: P2X 7 receptors were expressed by PCNA + proliferating cells. Spinal cord cross sections are shown (dorsal is up; asterisk indicates central canal; scale bar: 50 μm). Representative images of PCNA + and P2X 7 labelling in naïve fish (A-C′), at 7 dpi rostral (D-F′), and at 7 dpi caudal (G-I′). Colocalization (arrows) was seen in some cells (C′,F′,I′). PCNA + cells that did not colocalize with P2X 7 were also detected (open arrowheads) (C′,F′,I′). n =3/group. Created with BioRender.com.

Article Snippet: This included a P2X 7 control antigen (Alomone Labs, BLP-PR004, Q64663).

Techniques:

Journal: Biology Open

Article Title: P2X 7 regulates ependymo-radial glial cell proliferation in adult Danio rerio following spinal cord injury

doi: 10.1242/bio.060270

Figure Lengend Snippet: P2X 7 receptors were expressed by HuC/D + neurons. Spinal cord cross sections are shown (dorsal is up; asterisk indicates central canal; scale bar: 50 μm). Representative images of HuC/D + and P2X 7 labelling in naïve fish (A-C′), at 14 dpi rostral (D-F′), and at 14 dpi caudal (G-I′). Colocalization (arrows) was seen in larger mature neurons and smaller immature neurons (C′,F′,I′). n =3/group. Created with BioRender.com.

Article Snippet: This included a P2X 7 control antigen (Alomone Labs, BLP-PR004, Q64663).

Techniques:

Journal: Biology Open

Article Title: P2X 7 regulates ependymo-radial glial cell proliferation in adult Danio rerio following spinal cord injury

doi: 10.1242/bio.060270

Figure Lengend Snippet: Temporal changes in P2X 7 protein expression following SCI in adult zebrafish. Schematic representations of naïve and injured spinal cord areas of analysis (A-B), as well as tissue collection timeline (C). Representative western blot and corresponding total protein in naïve zebrafish (Zf) spinal cord tissue, mouse (Ms) hippocampal tissue as a positive control (+), and pre-absorption of naïve zebrafish spinal cord tissue with a blocking peptide as a negative control (-) (D). Representative western blot and corresponding total protein at 1 dpi (E), 7 dpi (G), and 14 dpi (I). Quantitative analysis of the 50 kDa isoform showed significant downregulation within the injury (In, n =10, one-way ANOVA with Dunnett's post hoc test, P =0.0079) and caudal to the lesion (Ca, n =9, P =0.0046) at 7 dpi when normalized to naïve tissue ( n =9) (H). Protein expression of the 50 kDa isoform showed basal levels of expression at 1 dpi (F) and 14 dpi (J). Data presented as means±s.d. **, significant differences, P <0.01. Created with BioRender.com.

Article Snippet: This included a P2X 7 control antigen (Alomone Labs, BLP-PR004, Q64663).

Techniques: Expressing, Western Blot, Positive Control, Blocking Assay, Negative Control

Journal: bioRxiv

Article Title: Synthesis, anticancer properties, and biological profiling of synthetic glycan analogs of proscillaridin A

doi: 10.1101/2025.03.31.646140

Figure Lengend Snippet: Flow cytometry profiling of key markers and cell cycle analysis in human and murine colon cancer cells. A) HCT-116 cells treated with 1 μM of compound 5 exhibit significantly enhanced surface expression of cleaved caspase-3, the hallmark of pre-apoptotic cell death. B) Effect of compounds 1 through 5 on surface CD133 (prominin 1) expression. C) Effect of compounds 1 through 5 on surface CD117 (c-Kit) expression. D) Cell cycle analysis by flow cytometry on HCT-116, HT-29, and CT26 cells treated with 100 nM of compounds 1 through 5

Article Snippet: Cleaved caspase-3 anti-mouse monoclonal antibody (Cat. #Ab156544) and CD117/c-Kit anti-mouse monoclonal antibody (Cat. #Ab096639) were obtained from Aladdin Scientific Corp. Anti-CD133 monoclonal antibody HB#7 was obtained from the DSHB and deposited there by Swaminathan, S.K./Panyam, J./Ohlfest, J.R. Ribonuclease A was obtained from Research Products International (97% purity).

Techniques: Flow Cytometry, Cell Cycle Assay, Expressing

Journal: Life Science Alliance

Article Title: Leishmania -infected macrophages release extracellular vesicles that can promote lesion development

doi: 10.26508/lsa.202000742

Figure Lengend Snippet: (A) Workflow for collection of extracellular vesicles from RAW264.7 macrophages infected with L. donovani parasites. Infection was performed in media supplemented with exosome-depleted serum. After 24 h, the culture medium was removed. Cultures were washed to remove uninternalized parasites and replenished with fresh medium supplemented with exosome-depleted serum. After an additional 48 h, culture medium was recovered, pooled, and processed following the centrifugation and filtration steps shown in the figure. (B) Nanoparticle tracking analysis was performed from which particle size distribution and particle concentration was obtained. Plot of particles/cell was calculated using cell count at the end of the infection. Data for graphs were obtained from multiple experiments (ceEV n = 7, LieEV n = 7; * P = 0.0082). (C) Representative image of vesicles in LieEV preparation processed for scanning electron microscopy. (D) Representative transmission electron microscopy image of immunogold CD9-labeled particles in LieEVs. Arrows point to gold particles denoting reactivity of antibody.

Article Snippet: The HM20-embedded samples were sliced into 100 nm thin sections that were placed on nickel grids, which were then immunogold-labeled with an anti-CD9 antibody (1/50 dilution vol/vol) (PB9930; Boster Bio) using a previously described method ( ).

Techniques: Infection, Centrifugation, Filtration, Concentration Assay, Cell Counting, Electron Microscopy, Transmission Assay, Labeling

Journal: Life Science Alliance

Article Title: Leishmania -infected macrophages release extracellular vesicles that can promote lesion development

doi: 10.26508/lsa.202000742

Figure Lengend Snippet: The proteome of LieEVs recovered after 72-h infection was compared with the proteome of ceEVs from uninfected cells. (A) Venn diagram was plotted using proteins identified by mass spectrometry and partitioned according to sample type. The presence and absence of host molecules with and without infection and common to both samples are indicated. (B) The protein content of the preparations was revealed by Ponceau S staining of the blots to normalize for material loaded into each well for each sample type. Approximately 1 × 10 10 particles from EV preparations from three replicate experiments and 50 μg of lysates from infected cells at the 72 h infection point were analyzed. (C) Western blot was performed to confirm the presence of known and novel exosome markers. Uninfected macrophages were treated in an identical manner as infected samples. The blots were then probed with anti-CD9, stripped, and probed with anti-Annexin A3. Identical blots were probed initially with anti-CD63, stripped, and probed with anti-calnexin. (D) Quantification was performed by measuring the mean gray background area using ImageJ software. Background pixel density was subtracted from the inverse of each measurement to obtain relative quantification values. Analysis of the blots showed that CD9 was significantly more abundant in LieEVs than ceEVs and cell lysates (* P = 0.0261, n = 3), whereas levels of CD63 were comparable for all samples. Annexin A3 was significantly more abundant in LieEVs than in ceEVs (* P = 0.0123, n = 3). Calnexin was barely detected in EVs as compared with cell lysates. Statistical test for differences was by ANOVA. Source data are available for this figure.

Article Snippet: The HM20-embedded samples were sliced into 100 nm thin sections that were placed on nickel grids, which were then immunogold-labeled with an anti-CD9 antibody (1/50 dilution vol/vol) (PB9930; Boster Bio) using a previously described method ( ).

Techniques: Infection, Mass Spectrometry, Staining, Western Blot, Software, Quantitative Proteomics

Journal: Bioactive Materials

Article Title: Bioengineered platelet nanoplatform enables renal-targeted dexamethasone delivery for chronic nephritis therapy with dual anti-inflammatory/anti-fibrotic effects and minimized systemic toxicity

doi: 10.1016/j.bioactmat.2025.06.002

Figure Lengend Snippet: LN-DEX@PLT ameliorates renal inflammation and reduce podocytes senescence in AD mice. (A–D) Expression of IL-6, TNF-α, FN, Col-I, in the kidney tissue 21 days post-treatment. (E) Schematic diagram of LN-DEX@PLT ameliorates renal inflammation and reduce podocytes senescence. (F) p53 value in the kidney tissue. (G) Immunofluorescence staining of p21. (H) CDK2 value in the kidney tissue. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: Additionally, anti-p53 DINP1, anti-F4/80/ADGRE1, and anti-CDK2 antibodies were procured from Boster Biological Technology, USA.

Techniques: Expressing, Immunofluorescence, Staining