pkd2l1  (Proteintech)

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 1 Pkd2l1+ CSF-cNs have the characteristics of both immature neurons and adult NSCs. a Immunofluorescence staining showing expression of the NSC marker Nestin in Pkd2l1+ CSF-cNs. b Pkd2l1+ CSF-cNs expressed the NSC marker Sox2 by immunofluorescence staining. c Pkd2l1+ CSF-cNs expressed the NSC marker CD133 by immunofluorescence staining. d Pkd2l1+ CSF-cNs did not express the NSC marker GFAP by immunofluorescence staining. e Pkd2l1+ CSF-cNs expressed the neuroblast marker doublecortin (Dcx) by immunofluorescence staining. f Immunofluorescence staining showed that Pkd2l1+ CSF-cNs expressed the immature neuronal marker βIII-tubulin (red). Scale bars, 20 µm. Nuclear counterstaining, DAPI.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining, Expressing, Marker

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 2 Pkd2l1 deletion results in the loss of CSF-cNs. a Schematic showing the generation and identification of genetically homogeneous Pkd2l1−/−mice. b, c Immunofluorescence staining of spinal cord sections showed that Pkd2l1 deletion caused the obviously reduced proliferation of GABA-positive cells after SCI (GABA: CSF-cNs marker) (n = 6). d, e The depletion of Pkd2l1 caused a significant reduction in Gata2-positive cells after SCI, as shown by immunofluorescence staining (Gata2: CSF-cNs marker) (n = 6). f, g Immunofluorescence staining shows that Pkd2l1 deletion resulted in almost the complete absence of Gata3-positive cell proliferation after SCI (Gata3: CSF-cNs marker) (n = 6). ***p < 0.001. Data are presented as the mean ± SD. Scale bars, 50 µm. Nuclear counterstaining, DAPI.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining, Marker

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 3 Pkd2l1 deletion suppresses the proliferation of CSF-cNs and leads to the loss of migratory CSF-cNs following SCI. a, b The EdU proliferation assay revealed a significant reduction in EdU-positive cells in Pkd2l1−/−mice compared to wild-type counterparts (n = 6). c, d Immunofluorescence analysis revealed a notable decrease in Dcx-positive cells following SCI in Pkd2l1−/−mice compared to their wild- type counterparts (n = 6). *p < 0.05, **p < 0.01. Data are presented as the mean ± SD. Scale bars, 50 µm.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Proliferation Assay

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 4 Pkd2l1 deletion retards the self-renewal of NSCs surrounding the central canal of the spinal cord after SCI. a, b Immunofluorescence staining showed that Pkd2l1−/−mice had almost no Nestin-positive cells surrounding the central canal of the spinal cord after SCI (n = 6). c, d Sox2 expression evidently decreased in Pkd2l1−/−mice after SCI, as shown by immunofluorescence staining (n = 6). e, f GFAP expression was reduced in Pkd2l1−/−mice after SCI, as shown by immunofluorescence staining (n = 6). ***p < 0.001. Data are presented as the mean ± SD. Scale bars, 50 µm. Nuclear counterstaining, DAPI.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining, Expressing

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 5 Pkd2l1 deletion retards myelin and neuronal regeneration following SCI. a, b Luxol Fast Blue staining showed that myelin regeneration was almost depleted in Pkd2l1−/−mice (n = 6). c, d Nissl staining showed that Pkd2l1−/−mice had a notable reduction in Nissl bodies surrounding the central canal following SCI (n = 6). *p < 0.05, **p < 0.01, ***p < 0.001. Data are presented as the mean ± SD: scale bars, 100 µm.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 7 Pkd2l1−/−mice restrained motor function recovery following SCI. a Measurement of BMS scores showed that the knockout of Pkd2l1 restricted motor function recovery in mice after SCI. b The loss of motor function in Pkd2l1−/−mice was evaluated based on the status of the hind limbs on day 35 following SCI (n = 6). *p < 0.05. compared to the Pkd2l1−/−SCI group. Data are presented as the mean ± SD.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Knock-Out

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 6 Pkd2l1 deletion promotes pericentral canal vacuolar degeneration and glial scar formation following SCI. a H&E staining showed that Pkd2l1−/−mice had more pericentral canal vacuolar degeneration after SCI. b, c Masson staining conducted on the transverse section of spinal cord tissue at the thoracic 10 (T10) level revealed that Pkd2l1−/−mice exhibited pronounced accumulation of collagen fibers, leading to the formation of glial scars around the central canal following SCI (n = 6). *p < 0.05. Data are presented as the mean ± SD. Scale bars, 100 µm.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 1 Pkd2l1+ CSF-cNs have the characteristics of both immature neurons and adult NSCs. a Immunofluorescence staining showing expression of the NSC marker Nestin in Pkd2l1+ CSF-cNs. b Pkd2l1+ CSF-cNs expressed the NSC marker Sox2 by immunofluorescence staining. c Pkd2l1+ CSF-cNs expressed the NSC marker CD133 by immunofluorescence staining. d Pkd2l1+ CSF-cNs did not express the NSC marker GFAP by immunofluorescence staining. e Pkd2l1+ CSF-cNs expressed the neuroblast marker doublecortin (Dcx) by immunofluorescence staining. f Immunofluorescence staining showed that Pkd2l1+ CSF-cNs expressed the immature neuronal marker βIII-tubulin (red). Scale bars, 20 µm. Nuclear counterstaining, DAPI.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining, Expressing, Marker

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 2 Pkd2l1 deletion results in the loss of CSF-cNs. a Schematic showing the generation and identification of genetically homogeneous Pkd2l1−/−mice. b, c Immunofluorescence staining of spinal cord sections showed that Pkd2l1 deletion caused the obviously reduced proliferation of GABA-positive cells after SCI (GABA: CSF-cNs marker) (n = 6). d, e The depletion of Pkd2l1 caused a significant reduction in Gata2-positive cells after SCI, as shown by immunofluorescence staining (Gata2: CSF-cNs marker) (n = 6). f, g Immunofluorescence staining shows that Pkd2l1 deletion resulted in almost the complete absence of Gata3-positive cell proliferation after SCI (Gata3: CSF-cNs marker) (n = 6). ***p < 0.001. Data are presented as the mean ± SD. Scale bars, 50 µm. Nuclear counterstaining, DAPI.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining, Marker

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 3 Pkd2l1 deletion suppresses the proliferation of CSF-cNs and leads to the loss of migratory CSF-cNs following SCI. a, b The EdU proliferation assay revealed a significant reduction in EdU-positive cells in Pkd2l1−/−mice compared to wild-type counterparts (n = 6). c, d Immunofluorescence analysis revealed a notable decrease in Dcx-positive cells following SCI in Pkd2l1−/−mice compared to their wild- type counterparts (n = 6). *p < 0.05, **p < 0.01. Data are presented as the mean ± SD. Scale bars, 50 µm.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Proliferation Assay

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 4 Pkd2l1 deletion retards the self-renewal of NSCs surrounding the central canal of the spinal cord after SCI. a, b Immunofluorescence staining showed that Pkd2l1−/−mice had almost no Nestin-positive cells surrounding the central canal of the spinal cord after SCI (n = 6). c, d Sox2 expression evidently decreased in Pkd2l1−/−mice after SCI, as shown by immunofluorescence staining (n = 6). e, f GFAP expression was reduced in Pkd2l1−/−mice after SCI, as shown by immunofluorescence staining (n = 6). ***p < 0.001. Data are presented as the mean ± SD. Scale bars, 50 µm. Nuclear counterstaining, DAPI.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining, Expressing

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 5 Pkd2l1 deletion retards myelin and neuronal regeneration following SCI. a, b Luxol Fast Blue staining showed that myelin regeneration was almost depleted in Pkd2l1−/−mice (n = 6). c, d Nissl staining showed that Pkd2l1−/−mice had a notable reduction in Nissl bodies surrounding the central canal following SCI (n = 6). *p < 0.05, **p < 0.01, ***p < 0.001. Data are presented as the mean ± SD: scale bars, 100 µm.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 7 Pkd2l1−/−mice restrained motor function recovery following SCI. a Measurement of BMS scores showed that the knockout of Pkd2l1 restricted motor function recovery in mice after SCI. b The loss of motor function in Pkd2l1−/−mice was evaluated based on the status of the hind limbs on day 35 following SCI (n = 6). *p < 0.05. compared to the Pkd2l1−/−SCI group. Data are presented as the mean ± SD.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Knock-Out

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 6 Pkd2l1 deletion promotes pericentral canal vacuolar degeneration and glial scar formation following SCI. a H&E staining showed that Pkd2l1−/−mice had more pericentral canal vacuolar degeneration after SCI. b, c Masson staining conducted on the transverse section of spinal cord tissue at the thoracic 10 (T10) level revealed that Pkd2l1−/−mice exhibited pronounced accumulation of collagen fibers, leading to the formation of glial scars around the central canal following SCI (n = 6). *p < 0.05. Data are presented as the mean ± SD. Scale bars, 100 µm.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cerebrospinal fluid-contacting neurons: a promising source for adult neural stem cell transplantation in spinal cord injury treatment

doi: 10.3389/fcell.2025.1549194

Figure Lengend Snippet: Screening and Labeling of CSF-cNs by Pkd2l1 (A) Screening process for high-purity CSF-cNs. Primary CSF-cNs were extracted from the cervical medullary regions of neonatal mice within 24 h of birth. Lentiral particles were to label CSF-cNs. Successfully labeled CSF-cNs screened using puromycin. The CSF-cNs expressed green fluorescent protein (GFP). (B) After transfecting primary cells containing CSF-cNs with lentivirus, puromycin (1 μg/mL) was added to select CSF-cNs. After 72 h, only a small number of adherent cells remained viable. (C) CSF-cNs that were successfully labeled by lentiviral particles could adhere to the wall and continue to grow. GFP expression was observed using inverted microscopy. BF = Bright Field. (D) Neurospheres formed by the suspension growth of CSF-cNs (GFP + cells) in a single cell state. BF = Bright Field. (E) The results of cellular immunofluorescence showed that CSF-cNs (GFP + cells) were co-labeled with their specific marker, Pkd2l1. (F) Quantitative analysis of CSF-cNs (GFP + cells) co-labeled with Pkd2l1 revealed that nearly 100% (99.8%) of the GFP + cells were indeed Pkd2l1 + CSF-cNs.

Article Snippet: Primary cells were allowed to adhere for 1 h, after which lentivirus (Lentivirus-Pkd2l1-GFP-puromycin, Shanghai Genechem Co., Ltd.) was added to the medium for transfection over 24 h. Subsequently, the medium was replaced with fresh serum-free neural stem cell medium, and puromycin (2 g/mL, 3 μL) was added to select cells for 72 h. The surviving cells after selection were collected, adjusted to a cell density of 2 × 10^5/well, resuspended in fresh serum-free neural stem cell medium, and seeded into ultra-low attachment 6-well plates (Corning Inc., United States of America) for suspension culture in an incubator at 37°C with 5% CO2.

Techniques: Labeling, Expressing, Inverted Microscopy, Suspension, Immunofluorescence, Marker

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cerebrospinal fluid-contacting neurons: a promising source for adult neural stem cell transplantation in spinal cord injury treatment

doi: 10.3389/fcell.2025.1549194

Figure Lengend Snippet: Screening and Labeling of CSF-cNs by Pkd2l1 (A) Screening process for high-purity CSF-cNs. Primary CSF-cNs were extracted from the cervical medullary regions of neonatal mice within 24 h of birth. Lentiral particles were to label CSF-cNs. Successfully labeled CSF-cNs screened using puromycin. The CSF-cNs expressed green fluorescent protein (GFP). (B) After transfecting primary cells containing CSF-cNs with lentivirus, puromycin (1 μg/mL) was added to select CSF-cNs. After 72 h, only a small number of adherent cells remained viable. (C) CSF-cNs that were successfully labeled by lentiviral particles could adhere to the wall and continue to grow. GFP expression was observed using inverted microscopy. BF = Bright Field. (D) Neurospheres formed by the suspension growth of CSF-cNs (GFP + cells) in a single cell state. BF = Bright Field. (E) The results of cellular immunofluorescence showed that CSF-cNs (GFP + cells) were co-labeled with their specific marker, Pkd2l1. (F) Quantitative analysis of CSF-cNs (GFP + cells) co-labeled with Pkd2l1 revealed that nearly 100% (99.8%) of the GFP + cells were indeed Pkd2l1 + CSF-cNs.

Article Snippet: After deparaffinization, the tissue was permeabilized using a solution containing 0.25% Triton X-100, followed by blocking with a goat serum solution for 1 h. The primary antibodies used in this study included Pkd2l1 (AB9084, 1:700, Merck Millipore, MA, United States), Nestin (sc23927, 1:200, Santa Cruz Biotechnology, Dallas, TX, United States), Sox2 (1:400, CST, Danvers, MA, United States), GFAP (1:300, CST, Danvers, MA, United States), CD133 (66666-1-lg, 1:400, Proteintech, United States), EGFR (66455-1-lg, 1:500, Proteintech, United States), PCNA (10205-2-AP, 1:300, Proteintech, United States), NeuN (MAB377, 1:400, Millipore, United States), O4 (MAB1326, 1:400, RD, United States), β III tubulin (66240-1-Ig, 1:200, Proteintech, United States), S100β (66616-1-lg, 1:500, Proteintech, United States), and GABA (A2052, 1:200, Millipore, United States).

Techniques: Labeling, Expressing, Inverted Microscopy, Suspension, Immunofluorescence, Marker

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cerebrospinal fluid-contacting neurons: a promising source for adult neural stem cell transplantation in spinal cord injury treatment

doi: 10.3389/fcell.2025.1549194

Figure Lengend Snippet: CSF-cNs can be continuously passaged in vitro (A) Bright-field images of CSF-cNs suspension culture in vitro under an inverted microscope. CSF-cNs possess an excellent spheroid-forming ability in vitro . CSF-cNs in the unicellular state can form a large number of neurospheres with a diameter of 150–200 μm in 3–4 days and can be continuously passaged up to a maximum of 60 generations in vitro . (B) Cellular immunofluorescence images showing neurospheres formed by CSF-cNs (GFP + cells) co-labeled with their specific marker Pkd2l1. (C, D) Cellular immunofluorescence images showing that neurospheres formed by CSF-cNs (GFP + cells) expressed the neural stem cell markers Nestin, Sox2. Nestin is expressed in the cell membrane, Sox2 is expressed in the nucleus. Nuclei are counterstained with DAPI in all panels.

Article Snippet: After deparaffinization, the tissue was permeabilized using a solution containing 0.25% Triton X-100, followed by blocking with a goat serum solution for 1 h. The primary antibodies used in this study included Pkd2l1 (AB9084, 1:700, Merck Millipore, MA, United States), Nestin (sc23927, 1:200, Santa Cruz Biotechnology, Dallas, TX, United States), Sox2 (1:400, CST, Danvers, MA, United States), GFAP (1:300, CST, Danvers, MA, United States), CD133 (66666-1-lg, 1:400, Proteintech, United States), EGFR (66455-1-lg, 1:500, Proteintech, United States), PCNA (10205-2-AP, 1:300, Proteintech, United States), NeuN (MAB377, 1:400, Millipore, United States), O4 (MAB1326, 1:400, RD, United States), β III tubulin (66240-1-Ig, 1:200, Proteintech, United States), S100β (66616-1-lg, 1:500, Proteintech, United States), and GABA (A2052, 1:200, Millipore, United States).

Techniques: In Vitro, Suspension, Inverted Microscopy, Immunofluorescence, Labeling, Marker, Membrane

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