Journal: iScience

Article Title: The Drosophila connectome reveals axo-axonic synapses on descending neurons

doi: 10.1016/j.isci.2026.115624

Figure Lengend Snippet: Axcs make cholinergic axo-axonic synapses on the GFs as predicted by the MANC connectome (A) AN08B098-type neurons (green) form axo-axonic connections at distinct locations along giant fiber (red, GF) axons. The scale bars shown are 20 μm. (B) The AN08B098-GF connections reconstructed in Neuroglancer (Neuroglancer scale 530.45 μm/vh) mirror the morphology seen with fluorescence in A . (C) We validated AN08B098-to-GF connectivity using anti-bruchpilot to stain for T-bars in active zones (white) along AN08B098-type neurons and colocalized the resulting fluorescence with the GFs (red), which were filled with tetramethylrhodamine. The scale bars shown are 20 μm. (D) Neuroglancer reveals presynaptic sites at similar locations seen in colocalized fluorescent image (Neuroglancer scale 530.45 μm/vh). (E and F) XY and XZ plane views of the preparation shown in (A) and (B). The zoomed-in 6 μm (scale bars 3 μm) inlays show AN08B098 forming a single synapse with the GF in (E). The yellow arrows detail the precise location AN08B098 forms the Brp-positive chemical synapse (white) to the GFs in (F). Scale bars shown are 5 μm. (G–O) EM images showing monosynaptic connections between single GF (green) and AN08B098 neurons identified by the following MANC id: (G and H) 21041, (I) 21589, (J) 23949, (K) 152261, (L) 16900, (M) 20444, (N) 22275, and (O) 24038. Pre-and postsynaptic sites are detected in EM slices using a 3D convolutional neural network to identify T-bars (cyan dots) and postsynaptic densities (PSDs, magenta dots). (P–S) We expressed anti-choline acetyltransferase (anti-ChAT) and anti-GFP in AN08B098 neurons. Anti-ChAT colocalizes to AN08B098 cells, with particularly bright staining in the cell bodies. This finding suggests acetylcholine synthesis is present within these cells, validating connectome transmitter predictions for AN08B098. Scale bars shown are (P) 20 μm and (Q–S) 5 μm.

Article Snippet: A mouse primary antibody against bruchpilot (1:50, DSHB, NC82) was used to label presynaptic chemical active zones and coupled to secondary Goat anti-mouse Alexa Fluor 647 (1:500, 115-605-003).

Techniques: Fluorescence, Staining

Journal: Bioactive Materials

Article Title: Dual-function thermoresponsive antibiotic-loaded hydrogel with antimicrobial and osteogenic properties for implant-related infection control

doi: 10.1016/j.bioactmat.2026.02.044

Figure Lengend Snippet: In vivo immunohistochemical analysis (n = 6). (A) Representative immunolabeling images of the Control, PNVCL, and PNVCL/TC 25 mg/mL groups showing osteopontin (OPN) and osteocalcin (OCN) expression (immunopositive cells indicated by black arrows). (B) Mean scores (0–3) ± standard deviation for OPN immunostaining. (C) Mean scores (0–3) ± standard deviation for OCN immunostaining. Bars indicate statistically significant differences between groups (p < 0.05; Tukey HSD test).

Article Snippet: Endogenous peroxidase activity was quenched by incubation with 3% hydrogen peroxide for 1 h, followed by blocking of nonspecific binding sites with 1% bovine serum albumin for 12 h. The sections were then incubated with goat anti-osteopontin and goat anti-osteocalcin primary antibodies (sc-21742 and sc-30044, respectively; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: In Vivo, Immunohistochemical staining, Immunolabeling, Control, Expressing, Standard Deviation, Immunostaining

Journal: Bioactive Materials

Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

doi: 10.1016/j.bioactmat.2026.01.043

Figure Lengend Snippet: Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: Primary antibodies against NLRP3 (cat. no. 15101) were obtained from Cell Signaling Technology (Beverly, Massachusetts, USA).

Techniques: In Vitro, Western Blot, Expressing, Control, Immunofluorescence, Fluorescence, Staining, Comparison

Journal: Bioactive Materials

Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

doi: 10.1016/j.bioactmat.2026.01.043

Figure Lengend Snippet: The composite hydrogel inhibits post-SCI pyroptosis in neurons. (A) Immunofluorescence images of residual neurons existing in the anterior horn of the spinal cord (scale bar, 500 μm and 200 μm). (B) Immunofluorescence image of Caspase-1 expression of neurons 3 days after SCI (scale bar, 20 μm). (C) Quantitative analysis of relative fluorescence intensity of Caspase-1 protein expression in neurons within the specified groups (n = 3). (D) Immunofluorescence image of GSDMD-N expression of neurons 3 days after SCI (scale bar, 20 μm). (E) Quantitative analysis of relative fluorescence intensity of GSDMD-N protein expression in neurons within the specified groups (n = 3). (F) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein 3 days after SCI, GAPDH was utilized as a loading control. (G) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: Primary antibodies against NLRP3 (cat. no. 15101) were obtained from Cell Signaling Technology (Beverly, Massachusetts, USA).

Techniques: Immunofluorescence, Expressing, Fluorescence, Western Blot, Control, Comparison

Journal: Bioactive Materials

Article Title: Precisely regulated physically-crosslinked carriers enable synergetic release of bioactive factors for MSC-mediated cartilage regeneration

doi: 10.1016/j.bioactmat.2026.01.009

Figure Lengend Snippet: Biocompatibility and Bioactivity of PSF and KSF in vivo . (a) Transwell Assay of MSCs after treated with PBS, MAP and PSF. Scale bar = 200 μm. (b) Wound Healing Assay of MSCs at 0h and 24h. Scale bar = 200 μm. (c) Immunofluorescent staining of cell pellets after 21 days of co-culture. Blue: DAPI; Green: ActinGreen; Red: Aggrecan. Scale bar = 200 μm. (d) Alcian blue staining of 2D cultured MSCs after 14 days. Scale bar = 200 μm. (e) Cell viability of MSCs at day 3 after co-culture. (f) The cell number of MSCs migrated from upper to lower chamber in Transwell assay. (g) The average distance MSCs migrated from injured margin in wound healing assay. (h–j) qRT-PCR of Col2a1 , Acan and Sox9 mRNA relative expression ratio compared with PBS group. ns: p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Immunofluorescent staining was performed with Aggrecan primary antibody (13880-1-AP, Proteintech, USA), ActinGreen ( R37110 , Thermo, USA) and DAPI (Solarbio, China) and observed with 3D reconstruction under high content imaging system (PerkinElmer, Operetta CLS, USA).

Techniques: In Vivo, Transwell Assay, Wound Healing Assay, Staining, Co-Culture Assay, Cell Culture, Quantitative RT-PCR, Expressing

Journal: Science Advances

Article Title: Glycerol-mediated nose-to-brain codelivery of anti–IL-17 and anti-CD73 antibodies enhances immunotherapy for melanoma brain metastases

doi: 10.1126/sciadv.adx7966

Figure Lengend Snippet: ( A and B ) Mice underwent brain fluorescence imaging at 0, 0.5, 1, 2, 4, 8, 12, 24, and 48 hours postintranasal delivery of Cy7-labeled a–IL-17 (Cy7–a–IL-17) and Cy5-labeled a-CD73 (Cy5–a-CD73). ( C and D ) Quantitative analysis of the fluorescence intensity for Cy7–a–IL-17 (C) and Cy5-a-CD73 (D) ( n = 3). ( E and F ) Quantitative analysis of the fluorescence intensity for Cy7–a–IL-17 (E) and Cy5–a-CD73 (F) in isolated brains at 2 hours ( n = 3). ( G ) Representative fluorescence microscopy images of brain sections 2 hours after intranasal delivery or intravenous injection of FITC–a–IL-17 (green) or Cy5–a-CD73 (red). Nuclei were counterstained with DAPI (blue). Scale bars, 20 μm. ( H to K ) Quantitative analysis of fluorescence intensity in the olfactory bulb (H), cortex (I), hippocampus (J), and cerebellum (K) ( n = 3). ( L ) Representative images of brain immunofluorescence were captured 2 hours after intranasal delivery or intravenous injection of FITC-IgG (green) to observe the distribution of antibodies in brain tissue sections. Blood vessels were stained with CD31 (red). Scale bars, 20 μm. All statistics are expressed as means ± SD. Statistical significance was assessed using one-way analysis of variance (ANOVA) with Fisher’s LSD test, where * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. ns, nonsignificant.

Article Snippet: For precise cellular localization, the sections were subjected to immunofluorescence staining: The vascular endothelium was labeled with an anti-CD31 primary antibody (catalog no. GB12063-100, Servicebio, Hubei, China), visualized with a Cy3-conjugated secondary antibody, and counterstained with DAPI for nuclei.

Techniques: Fluorescence, Imaging, Labeling, Isolation, Microscopy, Injection, Olfactory, Immunofluorescence, Staining