Journal: Neuron

Article Title: GPR37L1 identifies spinal cord astrocytes and protects neuropathic pain after nerve injury

doi: 10.1016/j.neuron.2025.01.012

Figure Lengend Snippet: (A) Schematic for snRNA-seq. Single cell nuclei were isolated from the SDH of naive mice. (B) UMAP plot of 17,886 nuclei identifies 30 clusters from 7 major cell types, including neurons, oligodendrocytes, OPCs (oligodendrocyte progenitor cells), astrocytes, microglia, endothelial cells, and mural cells. (C) The proportion of 7 different major cell types. (D) Violin plots showing the gene expression levels of 12 GPCRs in astrocyte subclusters. (E) UMAP plot showing Gpr37l1 expression. (F) RNA-scope images illustrate the co-localization of Gpr37l1 (in red) and Aldh1l1 (in green) within the SDH astrocytes of naïve mice. Scale = 50 μm (low-magnification); Scale = 5 μm (high-magnification). See also Figure S1 .

Article Snippet: The PVDF membranes were incubated overnight at 4°C with primary antibodies against GPR37L1 (rabbit, 1:200, AGR-050, alomone labs), GLT1 (rabbit, 1:1000, ab205247, Abcam), GAT3 (rabbit, 1:500, AGT-003, alomone labs), β-actin (rabbit, 1:1000, 20536–1-AP, Proteintech), Transferrin Receptor (mouse, 2 μg/ml, # 13–6800, Thermo Fisher), and GAPDH (1:5,000, rabbit, AP0063, BioWorld).

Techniques: Sequencing, Single Cell, Isolation, Gene Expression, Expressing, RNAscope

Journal: Neuron

Article Title: GPR37L1 identifies spinal cord astrocytes and protects neuropathic pain after nerve injury

doi: 10.1016/j.neuron.2025.01.012

Figure Lengend Snippet: (A-B) Mechanical (A) and cold sensitivity (B) tests reveal no difference between Gpr37l1 +/+ and Gpr37l1 −/− naïve mice. (C-D) Hargreaves (C) and hot plate (D) tests showing increased heat sensitivity to thermal stimulation in Gpr37l1 −/− naive mice. (E) Rotarod motor coordination assessment shows no difference between Gpr37l1 +/+ and Gpr37l1 −/− naïve mice. n = 12; pink dots represent female mice, and blue dots represent male mice. (F) Schematic representing the sciatic nerve crush surgery in Gpr37l1 +/+ and Gpr37l1 −/− mice. (G-H) Gpr37l1 −/− mice show rapid induction and prolongation of maintenance of mechanical and thermal hyperalgesia compared to Gpr37l1 +/+ mice after SNC injury. n = 10 in Gpr37l1 +/+ , n = 9 in Gpr37l1 −/− . (I) Schematic representation of the spared nerve injury (SNI) surgery. (J) Spontaneous pain scores in Gpr37l1 +/+ and Gpr37l1 −/− mice 3 days post SNI. (K) Facial grimace score showing spontaneous pain in Gpr37l1 +/+ and Gpr37l1 −/− mice 3 days post SNI. Data were expressed as mean ± SEM and statistically analyzed by unpaired t-tests (A, B, C, E, J, and K), one-way ANOVA followed by Tukey’s test (D), and two-way ANOVA followed by Sidak post-hoc test (G and H). *p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; n.s. no significance. See also Figure S2 .

Article Snippet: The PVDF membranes were incubated overnight at 4°C with primary antibodies against GPR37L1 (rabbit, 1:200, AGR-050, alomone labs), GLT1 (rabbit, 1:1000, ab205247, Abcam), GAT3 (rabbit, 1:500, AGT-003, alomone labs), β-actin (rabbit, 1:1000, 20536–1-AP, Proteintech), Transferrin Receptor (mouse, 2 μg/ml, # 13–6800, Thermo Fisher), and GAPDH (1:5,000, rabbit, AP0063, BioWorld).

Techniques: Sensitivity

Journal: Neuron

Article Title: GPR37L1 identifies spinal cord astrocytes and protects neuropathic pain after nerve injury

doi: 10.1016/j.neuron.2025.01.012

Figure Lengend Snippet: (A) Schematic representation of SNI surgery in Gpr37l1 +/+ and Gpr37l1 −/− mice. (B) Low magnification images depict GFAP and Iba1 staining in SDH of naïve Gpr37l1 +/+ and Gpr37l1 −/− mice (n = 4/group). Scale bar, 100 μm. (C-D) High magnification images showing reconstruction of microglia (C) and astrocytes (D). Scale bar, 10 μm. (E) Low magnification images of GFAP and Iba1 staining in SDH of Gpr37l1 +/+ and Gpr37l1 −/− mice at SNI 1 day (n = 4/group). Scale bar, 100 μm. (F-G) High magnification images showing reconstruction of microglia (F) and astrocyte (G) from the ipsilateral SDH at SNI 1-day. Scale bar, 10 μm. (H) Quantification of GFAP and Iba1 integrated density. (I-J) Sholl analysis from Iba1 (I) and GFAP (J) signals in SDH. n = 20 cells of 4 mice in each group. (K-L) Immunohistochemistry images show increase in pSTAT3 (Tyr705) in Gpr37l1 −/− but not Gpr37l1 +/+ mice at SNI 1-day. Scale bars, 100 μm (low magnification), 20 μm (high magnification). Data were expressed as mean ± SEM and statistically analyzed by two-way ANOVA followed by Sidak post-hoc test (H and L, n = 4/group). *p < 0.05, ** p < 0.01. n.s. no significance. See also Figure S3 .

Article Snippet: The PVDF membranes were incubated overnight at 4°C with primary antibodies against GPR37L1 (rabbit, 1:200, AGR-050, alomone labs), GLT1 (rabbit, 1:1000, ab205247, Abcam), GAT3 (rabbit, 1:500, AGT-003, alomone labs), β-actin (rabbit, 1:1000, 20536–1-AP, Proteintech), Transferrin Receptor (mouse, 2 μg/ml, # 13–6800, Thermo Fisher), and GAPDH (1:5,000, rabbit, AP0063, BioWorld).

Techniques: Staining, Analysis, Immunohistochemistry

Journal: Neuron

Article Title: GPR37L1 identifies spinal cord astrocytes and protects neuropathic pain after nerve injury

doi: 10.1016/j.neuron.2025.01.012

Figure Lengend Snippet: (A) Schematic for snRNA-seq from the SDH of Gpr37l1 +/+ and Gpr37l1 −/− mice with and without 1-day SNI. (B) Unbiased clustering of snRNA-seq data (left). The subcluster analysis of astrocytes illustrates 8266 nuclei from 4 groups, with a total of 107,542 nuclei, distinctly separating 28 astrocyte subclusters in UMAP (right). (C) SDH astrocyte UMAPs of Gpr37l1 +/+ and Gpr37l1 −/− mice with and without 1-day SNI. Right, proportion of astrocyte counts in total cell counts in SDH across 4 treatment groups. (D) The proportion of 28 astrocyte subclusters in SDH across 4 different groups. (E) Characterization of astrocyte subclusters by specific markers in WT mice subjected to SNI surgery (1-day). (F) The proportion of the subcluster 12 in SDH of WT-Naïve and WT-SNI-1 day groups. (G) The proportion of subclusters 3, 8, 12, 13, 16, and 21 across four different groups. (H) The volcano plot illustrates altered genes across all astrocyte subclusters, comparing KO-SNI and WT-SNI conditions. (I) The Gene Ontology (GO) enrichment analysis from the down-regulated genes in all astrocyte subclusters. (J) Volcano map of differentially expressed genes in astrocytes subclusters 3, 8, and 16. See also Figure S4 and Table S2 - S4 .

Article Snippet: The PVDF membranes were incubated overnight at 4°C with primary antibodies against GPR37L1 (rabbit, 1:200, AGR-050, alomone labs), GLT1 (rabbit, 1:1000, ab205247, Abcam), GAT3 (rabbit, 1:500, AGT-003, alomone labs), β-actin (rabbit, 1:1000, 20536–1-AP, Proteintech), Transferrin Receptor (mouse, 2 μg/ml, # 13–6800, Thermo Fisher), and GAPDH (1:5,000, rabbit, AP0063, BioWorld).

Techniques: Analysis

Journal: Neuron

Article Title: GPR37L1 identifies spinal cord astrocytes and protects neuropathic pain after nerve injury

doi: 10.1016/j.neuron.2025.01.012

Figure Lengend Snippet: (A) Schematic of intra-spinal cord injection of Gpr37l1 -targeting siRNA (si- Gpr37l1 ) and control siRNA (si-Con). (B) Western blot showing the downregulation of GPR37L1 in SDH by si-RNA treatment (n = 3/group). (C-D) Von Frey (C) and Hargreaves (D) tests show decreased paw withdrawal threshold and latency after intra-SDH injection of Gpr37l1 -si-RNA (n = 10/group). (E-F) Western blot analysis shows that siRNA-mediated knockdown of Gpr37l1 reduces the expression of GLT-1 (E) but not GAT-3 (F) in SDH. n = 5/group. (G-I) Whole-cell patch clamp recording in spinal cord slices. (G) Top, fluorescence image (left) and contrast images with an electrode (right) showing a Vglut2+ neuron. Scale bars, 40 μm. Bottom, sEPSCs traces. (H-I) Quantification of the frequency (H) and amplitude (I) of sEPSCs. n= 23 cells from 4 mice per group. (J-K) Astrogliosis in SDH following intra-spinal cord injection of si- Gpr37l1 and si-Con in Aldh1l1-EGFP mice. (J) Left, schematic of spinal injection. Right, IHC showing increased astrogliosis (GFAP staining) in the ipsilateral (Ipsi) side compared to the contralateral (Contra) side of SDH. Scale bars, 100 μm (top) and 20 μm (bottom). (K) Quantification of integrated fluorescence density of the images. n = 4/group. Data were expressed as mean ± SEM and statistically analyzed by unpaired t-test (B, E, F, H, and I), two-way ANOVA followed by Sidak post-hoc test (C and D), and one-way ANOVA followed by Tukey’s test (K). *p < 0.05, ** P < 0.01, ***p < 0.001. n.s. no significance. See also Figure S5 .

Article Snippet: The PVDF membranes were incubated overnight at 4°C with primary antibodies against GPR37L1 (rabbit, 1:200, AGR-050, alomone labs), GLT1 (rabbit, 1:1000, ab205247, Abcam), GAT3 (rabbit, 1:500, AGT-003, alomone labs), β-actin (rabbit, 1:1000, 20536–1-AP, Proteintech), Transferrin Receptor (mouse, 2 μg/ml, # 13–6800, Thermo Fisher), and GAPDH (1:5,000, rabbit, AP0063, BioWorld).

Techniques: Knockdown, Injection, Control, Western Blot, Analysis, Expressing, Patch Clamp, Fluorescence, Staining

Journal: Neuron

Article Title: GPR37L1 identifies spinal cord astrocytes and protects neuropathic pain after nerve injury

doi: 10.1016/j.neuron.2025.01.012

Figure Lengend Snippet: (A) Schematic for proximity ligation assay (PLA) for GPR37L1 and GLT1 in cultured astrocytes. (B) Representative images of PLA signals (red) from astrocytes treated with vehicle (Veh) and maresin 1 (MaR1, 100 ng/ml, 15 min). Scale bar, 10 μm. (C) Quantification of PLA signals. 40 cells from 3 independent experiments were included. (D) Stimulated emission depletion (STED) image, enlarged from the box in the left panel, shows the proximity of GPR37L1 and GLT-1 within GFAP+ astrocytic processes, as indicated by white arrowheads. Scale bars, 2 μm (left) and 400 nm (right). (E) Diagram outlining experimental design of over-expression of Slc1a2 through virus injected in the ipsilateral SDH. (F) Immunofluorescence shows the presence of AAV-GFAP- Slc1a2 :overexpression-mCherry in astrocytes but not neurons on the ipsilateral SDH. Scale bars, 100 μm (top) and 20 μm (bottom). (G) AAV-GFAP- Slc1a2 :overexpression-mCherry is expressed in > 91% of astrocytes (256/281 cells from 3 mice), with > 94% specificity (241/256 cells from 3 mice). (H) Von Frey testing showing the prevention of SNI-induced mechanical allodynia following intra-spinal injection of AAV- Slc1a2 :OE in Gpr37l1 +/+ mice but not in Gpr37l1 −/− mice. n= 8/group. (I-J) Patch-clamp recording traces (I) and quantification (J) of sEPSCs in spinal cord slices. Notably, both the frequency and the amplitude of sEPSCs from Gpr37l1 +/+ mice are lower than those from Gpr37l1 −/− mice, subjected to GLT-1 overexpression (n= 17 cells from 4 mice per group). Data were expressed as mean ± SEM and statistically analyzed by unpaired t-test (C and J) and two-way ANOVA followed by Sidak post-hoc test (H). ** p < 0.01. See also Figure S6 .

Article Snippet: The PVDF membranes were incubated overnight at 4°C with primary antibodies against GPR37L1 (rabbit, 1:200, AGR-050, alomone labs), GLT1 (rabbit, 1:1000, ab205247, Abcam), GAT3 (rabbit, 1:500, AGT-003, alomone labs), β-actin (rabbit, 1:1000, 20536–1-AP, Proteintech), Transferrin Receptor (mouse, 2 μg/ml, # 13–6800, Thermo Fisher), and GAPDH (1:5,000, rabbit, AP0063, BioWorld).

Techniques: Transmission Assay, Proximity Ligation Assay, Cell Culture, Over Expression, Virus, Injection, Immunofluorescence, Patch Clamp

Journal: Neuron

Article Title: GPR37L1 identifies spinal cord astrocytes and protects neuropathic pain after nerve injury

doi: 10.1016/j.neuron.2025.01.012

Figure Lengend Snippet: (A) Diagram of experimental design and timeline of behavioral testing and sample collection following virus and tamoxifen injections. (B) AAV-mcherry fluorescence in transverse (top) and sagittal (bottom) spinal cord section, showing co-localization with GFAP-labeled astrocytes. Scale bars, 50 μm (top left), 200 μm (top right), and 400 μm (bottom). (C) Quantification of AAV[FLEXon]- Gpr37l1 :overexpression-mCherry, showing expression in > 91% of astrocytes (244/267 cells from 3 mice), with > 92% specificity (226/244 cells from 3 mice) in Aldh1l1-Cre mice. (D) Western blotting analysis shows increased GPR37L1 expression in SDH following AAV- Gpr37l1 treatment (n = 4/group). (E) Von Frey testing shows the prevention of SNI-induced mechanical allodynia by SDH overexpression of GPR37L1 in both male and female mice (n= 8/group). (F-G) Overexpression of GPR37L1 prevented astrogliosis (GFAP staining) but not microgliosis (Iba1 staining) in SDH of SNI mice. Scales bar, 100 μm. n = 4/group. (H-I) Patch-clamp recordings of sEPSCs in spinal cord slices, showing the traces (H) and quantification (I). Note that GPR37L1 overexpression inhibited the SNI-induced increases in the frequency and amplitude of sEPSCs. n= 8 cells of 3 mice in each group. (J) Diagram of SNI surgery and virus injections. (K) Von Frey testing showing the reversal of established mechanical allodynia by SDH overexpression of GPR37L1 in both male and female mice. n= 6/group. Data were expressed as mean ± SEM and statistically analyzed by unpaired t-test (D), two-way ANOVA followed by Sidak post-hoc test (E and K), and one-way ANOVA followed by Tukey’s test (G and I). *p < 0.05, ** p < 0.01, ***p < 0.001, ****p < 0.0001. n.s. no significance. See also Figure S7 .

Article Snippet: The PVDF membranes were incubated overnight at 4°C with primary antibodies against GPR37L1 (rabbit, 1:200, AGR-050, alomone labs), GLT1 (rabbit, 1:1000, ab205247, Abcam), GAT3 (rabbit, 1:500, AGT-003, alomone labs), β-actin (rabbit, 1:1000, 20536–1-AP, Proteintech), Transferrin Receptor (mouse, 2 μg/ml, # 13–6800, Thermo Fisher), and GAPDH (1:5,000, rabbit, AP0063, BioWorld).

Techniques: Over Expression, Virus, Fluorescence, Labeling, Expressing, Western Blot, Analysis, Staining, Patch Clamp

Journal: Neuron

Article Title: GPR37L1 identifies spinal cord astrocytes and protects neuropathic pain after nerve injury

doi: 10.1016/j.neuron.2025.01.012

Figure Lengend Snippet: (A) Intrathecal (i.t.) injection of MaR1 (100 ng), given two weeks after SNI, decreases mechanical allodynia in Gpr37l1 +/+ mice (n=8) but not Gpr37l1 −/− mice (n=8). (B) Perfusion of spinal cord slices with MaR1 (10 ng/ml) reduces the frequency and the amplitude of sEPSCs on Vglut2+ excitatory neurons in the SDH laminar Ⅱ 2 weeks after SNI. n=9 cells from 3 mice. (C) MaR1 treatment (100 ng/ml, 15 min) increases glutamate uptake in astrocytes. Note that inhibition of glutamate uptake by GLT-1 inhibitor Dihydrokainate acid (DHK, 1 mM) blocked the MaR1’s effect. (D) Diagram of intra-spinal microinjection of AAV- Gpr37l1 -mcherry in Gpr37l1 knock-out mice 7 days after SNI surgery. (E) Image of AAV-mcherry fluorescence (red) and anti-GPR37L1 staining (green) of the ipsisilateral and contralateral SDH following the AAV injection. Right, quantitation of fluorescence density. Scale bar, 100 μm. (F) Von Frey testing showing the reversal of established mechanical allodynia by SDH re-expression of GPR37L1 in Gpr37l1 knock-out mice. n= 8/group. (G-H) SDH re-expression of GPR37L1 inhibits astrogliosis (GFAP staining) but not microgliosis (Iba1 staining) in Gpr37l1 KO mice with SNI. Scale bar, 100 μm. n = 4/group. (I-J) Patch-clamp sEPSCs recordings in spinal cord slices. Note that GPR37L1 re-expression inhibited the SNI-induced sEPSC increases in frequency and amplitude. n= 24 cells of 4 mice in each group. (K) Schematic of experimental design and behavioral test following intrathecal injection of MaR1 (100 ng) in Gpr37l1 −/− mice treated with AAV- Gpr37l1 or control AAV. (L) MaR1 loses its analgesic action in Gpr37l1 −/− mice with SNI and control AAV treatment. (M) MaR1 exhibits analgesic action following GPR37L1 re-expression by SDH AAV- Gpr37l1 injection. Data were expressed as mean ± SEM and statistically analyzed by two-way ANOVA followed by Sidak post-hoc test (A and F), repeated measures one-way ANOVA followed by Tukey’s multiple comparisons tests (B), one-way ANOVA followed by Tukey’s test (B, C, L, and M), and unpaired t-test (E, H, and I). *p < 0.05, ** p < 0.01, ***p < 0.001, ****p < 0.0001. n.s. no significance. See also Figure S8 .

Article Snippet: The PVDF membranes were incubated overnight at 4°C with primary antibodies against GPR37L1 (rabbit, 1:200, AGR-050, alomone labs), GLT1 (rabbit, 1:1000, ab205247, Abcam), GAT3 (rabbit, 1:500, AGT-003, alomone labs), β-actin (rabbit, 1:1000, 20536–1-AP, Proteintech), Transferrin Receptor (mouse, 2 μg/ml, # 13–6800, Thermo Fisher), and GAPDH (1:5,000, rabbit, AP0063, BioWorld).

Techniques: Activation Assay, Activity Assay, Transmission Assay, Injection, Inhibition, Microinjection, Knock-Out, Fluorescence, Staining, Quantitation Assay, Expressing, Patch Clamp, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Prosaposin orchestrates a TGFβ1-driven paracrine loop between Schwann cells and gastric cancer to accelerate perineural invasion

doi: 10.1186/s13046-026-03652-3

Figure Lengend Snippet: Bidirectional PSAP-TGFβ1 Paracrine Signaling Sustains SCs–GC Crosstalk. A Schematic of paracrine communication between SCs and GC Cells. B The secretion of PSAP by GC cells increased after co-culture with SCs. C TGFβ1 expression was increased in SCs co-cultured with GC cells and treated with rPSAP. D TGFβ1 secretion was increased in SCs co-cultured with GC cells and treated with rPSAP. E – F The secretion and expression of PSAP in GC cells treated with rTGFβ1 were increased. G - H The TGFβ1 inhibitor (Asiaticoside) can inhibit the effect of rTGFβ1 on the secretion and expression of PSAP

Article Snippet: Secreted PSAP and TGFβ1 levels in cell supernatants were quantified using Human PSAP (CUSABIO, China, #CSB-E12837h) and Rat TGFβ1 (JIANGLAI BIO, China, # JL12342,) ELISA kits.

Techniques: Co-Culture Assay, Expressing, Cell Culture

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Prosaposin orchestrates a TGFβ1-driven paracrine loop between Schwann cells and gastric cancer to accelerate perineural invasion

doi: 10.1186/s13046-026-03652-3

Figure Lengend Snippet: PSAP Activates GPR37/RAC1/ACTB Signaling to Drive TGFβ1 Secretion in SCs. A Schematic representation of PSAP secreted by GC cells acting on SCs. B-D rPSAP enhances SCs proliferation, migration, invasion and clonogenic capacity. E Functional enrichment analysis showed that the cytoskeleton of SCs changed after co-culture significantly. F Phalloidin staining reveals cytoskeletal changes in SCs following co-culture and rPSAP treatment. G SPR demonstrated direct binding between PASP and GPR37. H SCs co-cultured and exposed to rPSAP significantly activated GPR37/RAC1/ACTB signaling axis and TGFβ1 expression. I-K GPR37 sustains RAC1–ACTB signaling to drive TGF-β1 production and SC aggressiveness, conferring responsiveness to rPSAP

Article Snippet: Secreted PSAP and TGFβ1 levels in cell supernatants were quantified using Human PSAP (CUSABIO, China, #CSB-E12837h) and Rat TGFβ1 (JIANGLAI BIO, China, # JL12342,) ELISA kits.

Techniques: Migration, Functional Assay, Co-Culture Assay, Staining, Binding Assay, Cell Culture, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Prosaposin orchestrates a TGFβ1-driven paracrine loop between Schwann cells and gastric cancer to accelerate perineural invasion

doi: 10.1186/s13046-026-03652-3

Figure Lengend Snippet: TGFβ1 Engages the TGFβ1/Smad4/Sortilin Axis to Amplify PSAP Secretion and PNI. A Schematic representation of TGFβ1 secreted by SCs acting on GC cells. B In vitro PNI assays showed that downregulation of SCs-derived TGF-β1 diminished GC cell PNI. C-E Asiaticoside can inhibit the effects of rTGFβ1 on the proliferation, invasion and PSAP secretion of GC cells. F Schematic representation of PSAP secretion. G TGFβ1/Smad4 signaling activation induces increased PSAP secretion through the suppression of Sortilin expression. H Dual-luciferase and ChIP-qPCR analyses showed that Smad4 binds the Sortilin promoter and represses its transcription, linking TGF-β1/Smad4 activation to Sortilin downregulation. I Sortilin overexpression suppresses PSAP secretion

Article Snippet: Secreted PSAP and TGFβ1 levels in cell supernatants were quantified using Human PSAP (CUSABIO, China, #CSB-E12837h) and Rat TGFβ1 (JIANGLAI BIO, China, # JL12342,) ELISA kits.

Techniques: In Vitro, Derivative Assay, Activation Assay, Expressing, Luciferase, ChIP-qPCR, Over Expression

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Prosaposin orchestrates a TGFβ1-driven paracrine loop between Schwann cells and gastric cancer to accelerate perineural invasion

doi: 10.1186/s13046-026-03652-3

Figure Lengend Snippet: Therapeutic Targeting of PSAP/TGFβ1 Suppresses PNI In Vivo. A Inhibition of PSAP/TGFβ1 reduces PNI Incidence DRG-PNI model in vitro . B Schematic of a Sciatic Nerve Invasion Murine Model. C-D Tumor formation and H&E and IHC detection in the sciatic nerve model. E – F Functional and physiological assessments post intervention (Sciatic Nerve Score, Body Weight). G GMulticolor IF was used to verify the spatial distribution of PSAP, TGFβ1 and SCs(S100β) in PNI-positive tumor tissues

Article Snippet: Secreted PSAP and TGFβ1 levels in cell supernatants were quantified using Human PSAP (CUSABIO, China, #CSB-E12837h) and Rat TGFβ1 (JIANGLAI BIO, China, # JL12342,) ELISA kits.

Techniques: In Vivo, Inhibition, In Vitro, Functional Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Prosaposin orchestrates a TGFβ1-driven paracrine loop between Schwann cells and gastric cancer to accelerate perineural invasion

doi: 10.1186/s13046-026-03652-3

Figure Lengend Snippet: PSAP/TGFβ1/S100β multimodal signatures correlate with PNI and improve PNI discrimination. A IHC Staining of PSAP, TGFβ1, and S100β in PNI Subgroups. B Kaplan–Meier Survival Analysis by PNI Status in a Clinical Cohort. C Nomogram of PSAP, TGFβ1 and S100β combined to predict PNI. D - F ROC curve analysis, DCA, and calibration curves for comparison of PTS model and Baseline model for predicting PNI status. G The PTS model performance SHAP analysis. H - I ROC and calibration curves for the PTS and baseline models in an independent validation cohort. J Reclassification analysis of the PTS model and Baseline model

Article Snippet: Secreted PSAP and TGFβ1 levels in cell supernatants were quantified using Human PSAP (CUSABIO, China, #CSB-E12837h) and Rat TGFβ1 (JIANGLAI BIO, China, # JL12342,) ELISA kits.

Techniques: Immunohistochemistry, Comparison, Biomarker Discovery

Journal: Journal of Molecular Neuroscience

Article Title: Prosaposin Reduces α-Synuclein in Cells and Saposin C Dislodges it from Glucosylceramide-enriched Lipid Membranes

doi: 10.1007/s12031-022-02066-y

Figure Lengend Snippet: Antibodies used for Western blot

Article Snippet: PSAP-GFP (HG16224-ACG, Sino Biological) and control-EGFP (13031, Addgene) plasmids underwent amplification in E. coli and were purified using plasmid extraction kits (12143, Qiagen).

Techniques: Western Blot

Journal: Journal of Molecular Neuroscience

Article Title: Prosaposin Reduces α-Synuclein in Cells and Saposin C Dislodges it from Glucosylceramide-enriched Lipid Membranes

doi: 10.1007/s12031-022-02066-y

Figure Lengend Snippet: Overexpression of Prosaposin (PSAP) lowers levels of α-synuclein. ( a ) Representative Western Blot for anti-GFP (Left) and anti-PSAP (Right) in EGFP-Overexpressing (EGFP-OE) cells and PSAP-Overexpressing (PSAP-OE) cells showing PSAP-GFP at 105 kDa, endogenous PSAP at 70 kDa, and control EGFP at 32 kDa. ( b ) Representative live-cell images of PSAP-OE cells for PSAP-GFP co-localized with the lysosome (Green: PSAP-GFP, Magenta: LysoTracker, scale bar 5 µm). ( c ) Representative Western Blot for anti-α-synuclein and anti-β-actin in EGFP-OE cells and PSAP-OE cells. ( d ) Quantification of Western Blot analysis showing levels of α-synuclein in EGFP-OE cells (n = 10) and PSAP-OE cells (n = 8). ( e ) Real-time PCR analysis showing mRNA levels of SNCA in EGFP-OE cells (n = 6) and PSAP-OE cells (n = 6). ( f ) Quantification of α-synuclein levels in cell culture media of EGFP-OE cells (n = 8) and PSAP-OE cells (n = 8) measured by enzyme-linked immunosorbent assay. ( g ) Representative Western blot for anti-Glucocerebrosidase (GCase) and anti-GAPDH in EGFP-OE cells and PSAP-OE cells. ( h ) Quantification of Western blot analysis showing levels of GCase in EGFP-OE cells (n = 8) and PSAP-OE cells (n = 8). ( i ) Non-specific GBA activity and specific GCase activity analysis in EGFP-OE cells (n = 8) and PSAP-OE cells (n = 8) treated with or without AMP-deoxynojirimycin. Data are expressed as mean ± SEM. n.s: non-significant, p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001, Mann–Whitney U test (d, e, f, h,) and Two-way ANOVA with Tukey’s multiple comparison test (i)

Article Snippet: PSAP-GFP (HG16224-ACG, Sino Biological) and control-EGFP (13031, Addgene) plasmids underwent amplification in E. coli and were purified using plasmid extraction kits (12143, Qiagen).

Techniques: Over Expression, Western Blot, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay, Activity Assay, MANN-WHITNEY