Journal: Cellular signalling

Article Title: Vegfr3 activation of Pkd2l1 + CSF-cNs triggers the neural stem cell response in spinal cord injury.

doi: 10.1016/j.cellsig.2025.111675

Figure Lengend Snippet: Fig. 1. Vegfr3 is expressed primarily in Pkd2l1+ CSF-cNs. (A) A diagram illustrating the process of isolating and culturing primary CSF-cNs. (B) Pkd2l1+ CSF-cNs expressed Vegfr1, Vegfr2, and Vegfr3, with Vegfr3 showing the highest expression in vitro. (C) Western blot analysis of Vegfr1, Vegfr2, and Vegfr3 in Pkd2l1+ CSF- cNs. (D) Strategy diagram for the construction of Pkd2l1-2 A-CreERT2 × R26-CAG-LSL-ZsGreen transgenic mice, outlining the genetic modifications and breeding strategy employed. (E) Immunofluorescence colabeling of Pkd2l1 and the inherent ZsGreen fluorescence in Pkd2l1-2 A-CreERT2 × R26-CAG-LSL-ZsGreen transgenic mice. The overlap of the Pkd2l1 and ZsGreen signals confirmed the successful generation of the transgenic model. (F) Coexpression analysis showing Vegfr3 localization in Pkd2l1+ CSF-cNs. Double immunofluorescence staining revealed the colocalization of Vegfr3 with Pkd2l1+ cells, indicating that Vegfr3 is expressed in these neurons. (G) Immunofluorescence images illustrate the lack of colocalization between Vegfr3 and Foxj1, a specific marker for ependymal cells, in both the sham and SCI 7 d groups. These findings indicate that Vegfr3+ cells are distinct from ependymal cells. (H) Immunofluorescence images show the absence of colocalization between Vegfr3 and ALDH1L1, an astrocyte marker, in both the sham and SCI 7 d groups. Scale bar: 50 μm (B, E, F, G, H).

Article Snippet: For mIHC, the following primary antibodies were used: Pkd2l1 (#AB9084, 1:700, Merck Millipore), CD133 (#66666–1-Ig, 1:500, Proteintech), GFAP (#3670, 1:300, Cell Signaling Technology), EGFR (#66863–1-lg, 1:500, Proteintech), and Nestin (Sc-6251, 1:300, Santa Cruz Biotechnology).

Techniques: Expressing, In Vitro, Western Blot, Transgenic Assay, Immunofluorescence, Fluorescence, Double Immunofluorescence Staining, Marker

Journal: Cellular signalling

Article Title: Vegfr3 activation of Pkd2l1 + CSF-cNs triggers the neural stem cell response in spinal cord injury.

doi: 10.1016/j.cellsig.2025.111675

Figure Lengend Snippet: Fig. 3. Role of Vegfr3 in the proliferation and activation of Pkd2l1+ CSF-cNs. (A, B) The overexpression and knockdown efficiencies of Vegfr3 in Pkd2l1+ CSF-cNs were confirmed through qRT-PCR and Western blotting following transfection. (C) Representative images of CSF-cNs and the average diameter of neurospheres with or without Vegfr3 overexpression or inhibition. (D) Representative immu nofluorescence images and quantification of EdU+ Pkd2l1+ CSF-cNs with or without Vegfr3 overexpression or inhibition. (E) Representative immunofluorescence images and quantification of pHH3+ Pkd2l1+ CSF-cNs with or without Vegfr3 overexpression or inhibition. (F) A CCK-8 assay was used to assess the viability of Pkd2l1+ CSF-cNs with either Vegfr3 overexpression or knockdown. (G) Representative Western blots showing the protein levels of EGFR, Ascl, and Sox2 in Pkd2l1+

Article Snippet: For mIHC, the following primary antibodies were used: Pkd2l1 (#AB9084, 1:700, Merck Millipore), CD133 (#66666–1-Ig, 1:500, Proteintech), GFAP (#3670, 1:300, Cell Signaling Technology), EGFR (#66863–1-lg, 1:500, Proteintech), and Nestin (Sc-6251, 1:300, Santa Cruz Biotechnology).

Techniques: Activation Assay, Over Expression, Knockdown, Quantitative RT-PCR, Western Blot, Transfection, Inhibition, Immunofluorescence, CCK-8 Assay