Journal: bioRxiv

Article Title: Adeno-associated viruses (AAVs) induce dose-dependent neonatal ventriculomegaly following intracerebroventricular administration

doi: 10.64898/2025.12.30.697113

Figure Lengend Snippet: AAVs induce dose-dependent CSF dyshomeostasis. (A) CSF CCL2 concentrations measured by ELISA at P7.5 following P0.5 ICV injection of escalating AAV doses. (B, C) Parenchymal microglial response. (B) Representative Iba1 immunostaining in P7.5 brain sections. (C) Quantification of Iba1⁺ cell density in parenchymal regions. (D, E) ChP microglial response. (D) Representative Iba1 immunostaining in the ChP. (E) Quantification of Iba1⁺ cell density in the ChP. (F) CSF transthyretin (TTR) concentrations measured by ELISA at P7.5 following P0.5 ICV injection of escalating AAV doses. (G–J) ChP protein analysis by immunoblot. (G) Representative blot for TTR and GFP in ChP lysates. (H) Relative GFP expression. (I) Relative TTR expression. (J) Correlation analysis between GFP and TTR levels in individual samples. (K, L) CSF ion disruption following inflammation. (K) Experimental timeline for LPS-induced inflammation and CSF collection in adult mice. (L) CSF potassium (K⁺) concentration measured by ICP-MS.

Article Snippet: The concentrations of C-C motif chemokine ligand 2 (CCL2) and transthyretin (TTR) in the CSF were quantified using commercial ELISA kits (mouse CCL2 ELISA kit, BSKM1011, Bioss; mouse TTR ELISA kit, ab282297, Abcam), according to the manufacturers’ protocols.

Techniques: Enzyme-linked Immunosorbent Assay, Injection, Immunostaining, Western Blot, Expressing, Disruption, Concentration Assay

Journal: bioRxiv

Article Title: Adeno-associated viruses (AAVs) induce dose-dependent neonatal ventriculomegaly following intracerebroventricular administration

doi: 10.64898/2025.12.30.697113

Figure Lengend Snippet: AAV induces dose-dependent embryonic and adult ventriculomegaly. (A) Experimental timeline for assessing acute AAV2/5 transfection 24 hours post in utero ICV injection. (B) Representative image confirming robust AAV2/5 transfection in the embryonic ChP at E14.5 (24 hours post-injection). (C) Timeline for evaluating the effects of escalating AAV2/5 doses via in utero ICV injection at E13.5, with analysis at E16.5. (D) Embryonic survival rates at E16.5. Numbers indicate the proportion of surviving embryos per litter. (E) Representative coronal sections of E16.5 embryonic brains showing transgene expression (GFP) and ventricular morphology. (F, G) CSF analysis from E16.5 embryos. (F) CCL2 concentrations. (G) Transthyretin (TTR) concentrations. (H) Timeline for assessing the effects of escalating AAV2/5 doses via ICV injection in adult mice (3-day endpoint). (I) Representative coronal sections at the level of the posterior anterior commissure (pAC; dashed outline) showing lateral ventricle morphology in adult brains. (J, K) Representative sagittal section demonstrating the differential cellular tropism of AAV2/5. (J) ChP-specific transfection by low-dose AAV2/5. (K) A more permissive tropism of high-dose AAV2/5, with ectopic GFP signal also observed in hippocampal CA1 region.

Article Snippet: The concentrations of C-C motif chemokine ligand 2 (CCL2) and transthyretin (TTR) in the CSF were quantified using commercial ELISA kits (mouse CCL2 ELISA kit, BSKM1011, Bioss; mouse TTR ELISA kit, ab282297, Abcam), according to the manufacturers’ protocols.

Techniques: Transfection, In Utero, Injection, Expressing

Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Exposed to High-Dose Ionizing Radiation Promote M2 Polarization of Macrophages, Which Induce Radiosensitivity in Cervical Cancer

doi: 10.3390/cancers15051620

Figure Lengend Snippet: CCL2 is critical for high-dose irradiated CAFs to promote M2 polarization. ( A ) mCAFs were exposed to 8 Gy radiation or not (control), and the supernatant of mCAFs was subjected to multiplex cytokine array analysis 24 h later. Left: a representative blot. Right: quantification of dot intensity of significantly changed cytokines. ( B ) Cervical cancer cell lines and CAFs were irradiated with the indicated doses. After 24 h, CCL2 expression was analyzed by Western blot analysis. ( C ) mCAFs and hCAFs were exposed to the indicated doses of radiation. After 24 h, the supernatant was subjected to ELISA. Data are shown as mean ± SD from three independent experiments. The data were analyzed using one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001. ( D ) BMDMs were treated with 20 ng/mL CCL2 for 24 h and then subjected to flow cytometry analysis. Data are shown as mean ± SD from three independent experiments. Differences between groups were analyzed using unpaired Student’s t -test. * p < 0.05. ( E ) M0 BMDMs treated with 10 nM CCR2 antagonist (INCB3344) were co-cultured with irradiated or non-irradiated mCAFs for 3 days. Then, the cells were subjected to flow cytometry analysis to evaluate CD206 expression. Data are shown as mean ± SD from three independent experiments. * p < 0.05. ( F ) Anti-CCL2 neutralizing antibody (20 μg/mL) was added to a co-culture system consisting of 8 Gy irradiated mCAFs and M0 BMDMs. The BMDMs were subjected to flow cytometry analysis after 3 days of co-cultivation. Data are shown as mean ± SD from three independent experiments. The data were analyzed using one-way ANOVA. * p < 0.05. ( G ) M0 BMDMs were co-cultured with mCAFs exposed to the indicated doses of radiation for 3 days and 10 nM CCR2 antagonist (INCB3344) was added into the 8 Gy irradiated mCAF co-culture system. Immunofluorescence analysis of the expression of Ym-1 (red) in BMDMs. The whole western blots are shown in File S1.

Article Snippet: The supernatant of CAFs and cervical cancer cells was assayed for cytokines and chemokines using the Mouse XL Cytokine Array (Ary028; R&D Systems) and CCL2 ELISA kits (mouse, EK0568; human, EK0441; BOSTER) according to the manufacturers’ instructions.

Techniques: Irradiation, Control, Multiplex Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cell Culture, Co-Culture Assay, Immunofluorescence

Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Exposed to High-Dose Ionizing Radiation Promote M2 Polarization of Macrophages, Which Induce Radiosensitivity in Cervical Cancer

doi: 10.3390/cancers15051620

Figure Lengend Snippet: Model summarizing the proposed signaling events among CAFs, TAMs, and cervical cancer cells under high-dose radiation. CAFs secrete an elevated level of CCL2 upon treatment with high-dose radiation. CCL2 promotes pro-tumor transition in macrophages. M2 macrophages induce radioresistance in cervical cancer cells.

Article Snippet: The supernatant of CAFs and cervical cancer cells was assayed for cytokines and chemokines using the Mouse XL Cytokine Array (Ary028; R&D Systems) and CCL2 ELISA kits (mouse, EK0568; human, EK0441; BOSTER) according to the manufacturers’ instructions.

Techniques: