Journal: Journal of Inflammation Research

Article Title: Electroacupuncture Inhibits the Early Neuroinflammatory Cascade Triggered by TLR2 in the Prodromal Period of PD

doi: 10.2147/JIR.S585729

Figure Lengend Snippet: EA can inhibit the activation of microglia in the pPD stage mediated by TLR2. ( A ) Western blot detection of TLR2 protein expression in the substantia nigra. ( B ) Western blot detection of MyD88 protein expression in the substantia nigra. ( C ) Western blot detection of p-NF-κB-p65 expression in the substantia nigra. ( D ) Western blot detection of NLRP3 protein expression in the substantia nigra. ( E ) Western blot detection of Caspase-1 protein expression in the substantia nigra. ( F ) Western blot detection of GSDMD protein expression in the substantia nigra. ( G ) Western blot detection of IL-1β protein expression in the substantia nigra. (n=6; compared with the control group, *** P <0.001; compared with the model group, ### P <0.001, ## P <0.01, # P <0.05).

Article Snippet: The membrane was incubated overnight at 4°C with primary antibodies specific to TLR2 (#bs-1019R, Bioss, 1:1000), MyD88 (#bs-1047R, Bioss, 1:1000), p-NF-κB-p65 (#bs-7343R, Bioss, 1:1000), NLRP3 (#bs-41293R, Bioss, 1:1000), caspase-1 (#bsm-52441R, Bioss, 1:1000), IL-1β (#bs-0812R, Bioss, 1:1000), and GSDMD (#bs-14287R, Bioss, 1:1000), followed by incubation with an IgG secondary antibody (#bs-10900R, Bioss, 1:5000) for 1 hour in the dark.

Techniques: Activation Assay, Western Blot, Expressing, Control

Journal: Journal of Inflammation Research

Article Title: Electroacupuncture Inhibits the Early Neuroinflammatory Cascade Triggered by TLR2 in the Prodromal Period of PD

doi: 10.2147/JIR.S585729

Figure Lengend Snippet: EA intervention affects the expression of TLR2 in different regions of pPD, inhibits microglial activation, regulates the TLR2/NF-κB/NLRP3 pathway, and suppresses pyroptosis of microglia.

Article Snippet: The membrane was incubated overnight at 4°C with primary antibodies specific to TLR2 (#bs-1019R, Bioss, 1:1000), MyD88 (#bs-1047R, Bioss, 1:1000), p-NF-κB-p65 (#bs-7343R, Bioss, 1:1000), NLRP3 (#bs-41293R, Bioss, 1:1000), caspase-1 (#bsm-52441R, Bioss, 1:1000), IL-1β (#bs-0812R, Bioss, 1:1000), and GSDMD (#bs-14287R, Bioss, 1:1000), followed by incubation with an IgG secondary antibody (#bs-10900R, Bioss, 1:5000) for 1 hour in the dark.

Techniques: Expressing, Activation Assay

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Outline of study time-course and sample processing for EV isolation. Antemortem plasma samples with postmortem pathological confirmation of neurological diagnoses were processed to isolate cell-specific EVs using our mTENPO microfluidic platform, alongside plasma protein biomarkers using commercial digital ELISA, for patients with LBD ( n = 30), AD ( n = 31), AD/LBD ( n = 30), AD/ALB ( n = 19), and controls ( n = 27). B The mTENPO platform, illustrating the external magnet, inlet reservoir, outlet ports, and tubing connections to syringe pumps. Syringes are connected to the waste outlet for blocking, washing, and sample addition steps, and then replaced and switched to the lysate outlet before captured EVs are lysed on-chip. The inset shows a photo of the mTENPO chip with a quarter for scale. C Schematic of operation of the mTENPO platform for cell-specific EV isolation using antibody-labeled magnetic nanoparticles (MNPs) for GluR2+ (top) and GLAST+ (bottom) EV pulldowns. D Scanning electron microscopy (SEM) images of GluR2+ EVs immobilized on the edges of pores of the mTENPO device’s surface. E SEM images of GLAST+ EVs immobilized on the edges of pores of the mTENPO device’s surface. F Representative cropped western blot images showing protein expression of GluR2, GLAST, and EV-associated marker TSG101 using mTENPO-isolated GluR2+ or GLAST + EV lysates from n = 2 human plasma samples. Full-length western blot images are shown in Supplementary Fig. .

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Isolation, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Blocking Assay, Labeling, Electron Microscopy, Western Blot, Expressing, Marker

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Heatmap of z-score of log 2 (expression) for biomarkers with Benjamini-Hochberg FDR-corrected P value < 0.1. Subjects (columns) are hierarchically clustered within cohort and biomarkers within each compartment (rows) are sorted by descending fold-change. B Volcano plot demonstrating differential expression of GluR2+ EV miRNAs, GLAST + EV miRNAs, and plasma proteins. C Venn diagram showing overlap in FDR P value significant miRNAs ( P value < 0.1) between GluR2+ EVs and GLAST+ EVs. D Top 30 biomarkers in all compartments ranked by descending area under the curve (AUC). Error bars represent standard error from bootstrapping 10x.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Expressing, Quantitative Proteomics, Clinical Proteomics

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: GO and KEGG pathway analyses were performed on differentially expressed miRNAs using DIANA miRPath v4.0 using the TarBase v8.0 database. FDR P values for identified GO terms and KEGG pathways were calculated using a one-sided Fisher’s exact test and considered significant at P value < 0.05. The top 10 (ranked by number of target genes) terms within each of the three GO categories (BP, CC, MF) and top 10 (ranked by number of target genes) KEGG pathways were identified for each pulldown. A Top 10 terms within each GO category for GluR2+ EV miRNAs. B Top 10 KEGG pathways for GluR2+ EV miRNAs. C Top 10 terms within each GO category for GLAST + EV miRNAs. D Top 10 KEGG pathways for GLAST + EV miRNAs. In all panels, each bar is labeled to the right with the number of differentially expressed miRNAs associated with the given GO term or KEGG pathway.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Labeling

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Heatmap of z-score of log 2 (expression) for LASSO-selected biomarkers. Subjects (columns) are hierarchically clustered within cohort and biomarkers within each compartment (rows) are sorted by descending AUC. B Kendall correlation staircase plots identifying the extent to which biomarker information was correlated between the LASSO-selected GluR2+ EV, GLAST + EV, and protein biomarkers. Biomarkers are sorted within compartments by AUC. The inset shows the correlation distribution of Kendall’s τ, where the dotted line represents the median count. C LASSO panel accuracy versus panel size for classifying LBD versus AD, shown in blue; accuracy is assessed through tenfold cross-validation, with error bars representing standard error from 5 repeats of panel training on the LBD vs AD patient groups. Average accuracy and standard error for control experiments performed by scrambling patient labels 10x are shown in orange. D LASSO panel AUC versus panel size for classifying LBD versus AD, shown in blue with error bars as described in ( C ). Average AUC and standard error for the same control experiments described in ( C ) are shown in orange. E AUCs for the 15-marker LASSO panel and individual LASSO biomarkers, sorted by descending AUC. Error bars represent 95% confidence intervals, calculated from 5x repeats of tenfold cross-validation for the 15-marker panel or from bootstrapping 10x for individual markers.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Expressing, Biomarker Discovery, Control, Marker

Journal: Molecular Cancer

Article Title: HnRPD/AUF1 facilitates human ovarian cancer progression through activating FLI1 and maintaining cisplatin resistance

doi: 10.1186/s12943-026-02599-5

Figure Lengend Snippet: HnRPD is identified to be an RAB8A mRNA-interacting protein. A The mRNA levels of RAB8A in OVCAR-8 cells transfected with a vector expressing GPR137 or a control empty vector (Con) and cultured with actinomycin D (AcD) for the indicated time periods. * p < 0.05; ** p < 0.01 versus Con; error bar, SD. N = 3. B qRT-PCR analysis of co-precipitated RAB8A mRNA by GPR137 antibody in the RIP assay in SK-OV-3 cells. IgG was used as a negative control. N = 3. C qRT-PCR analysis of co-precipitated RAB8A mRNA by GPR137 antibody in the RIP assay in A2780 cells. IgG was used as a negative control. N = 3. D An experimental model showing the procedure of IP and RNA pull-down with LC-MS/MS analysis. E LC-MS/MS analysis showing the identified proteins with difference in abundance from GPR137 antibody-IP group versus the corresponding control. F LC-MS/MS analysis showing the identified proteins with difference in abundance from RAB8A RNA pull-down group versus the corresponding control

Article Snippet: GPR137 (bs-16270R, 1:1000) antibody and RAB8A (bs-6176R, 1:1000) antibody were acquired from Bioss Biotechnology (Beijing, China), while Flag-tag (M185-3, 1:10000) antibody was obtained from MBL Biotechnology (Beijing, China).

Techniques: Transfection, Plasmid Preparation, Expressing, Control, Cell Culture, Quantitative RT-PCR, Negative Control, Liquid Chromatography with Mass Spectroscopy

Journal: Molecular Cancer

Article Title: HnRPD/AUF1 facilitates human ovarian cancer progression through activating FLI1 and maintaining cisplatin resistance

doi: 10.1186/s12943-026-02599-5

Figure Lengend Snippet: Molecular docking analysis of GPR137-hnRPD and hnRPD- RAB8A RNA. A Docking results by HDOCK server. Yellow indicates GPR137, purple indicates hnRPD. B Docking results visualized in the form of Surface. Red indicates protein-protein contact area. C Interaction of GPR137-hnRPD analyzed by PLIP. D Docking results by HDOCK server. Green indicates RAB8A RNA, purple indicates hnRPD. E Docking results visualized in the form of Surface. Red indicates protein-RNA contact area. F Interaction of hnRPD-RAB8A RNA analyzed by PLIP

Article Snippet: GPR137 (bs-16270R, 1:1000) antibody and RAB8A (bs-6176R, 1:1000) antibody were acquired from Bioss Biotechnology (Beijing, China), while Flag-tag (M185-3, 1:10000) antibody was obtained from MBL Biotechnology (Beijing, China).

Techniques:

Journal: Molecular Cancer

Article Title: HnRPD/AUF1 facilitates human ovarian cancer progression through activating FLI1 and maintaining cisplatin resistance

doi: 10.1186/s12943-026-02599-5

Figure Lengend Snippet: GPR137 promotes RAB8A mRNA stability through elevating hnRPD expression. A Co-IP of endogenous GPR137 and hnRPD in SK-OV-3 cells. IP: GPR137, WB: hnRPD. IgG was used as a negative control. B Co-IP of endogenous hnRPD and GPR137 in A2780 cells. IP: hnRPD, WB: GPR137. IgG was used as a negative control. C qRT-PCR analysis of co-precipitated RAB8A mRNA by hnRPD antibody in the RIP assay in SK-OV-3 cells. IgG was used as a negative control. ** p < 0.01 versus IgG; error bar, SD. N = 3. D RNA pull-down assay in A2780 cells with RAB8A RNA or control RNA fragments. Protein abundance was normalized to GAPDH. E The mRNA expression of hnRPD in SK-OV-3 cells transfected with GPR137 shRNA or scrambled shRNA (shRNA-Con). Error bar, SD. N = 3. F The mRNA expression of hnRPD in OVCAR-8 cells transfected with a vector expressing GPR137 or a control empty vector (Con). Error bar, SD. N = 3. G The protein expression of GPR137 and hnRPD in SK-OV-3 cells transfected with GPR137 shRNA (shGPR137, +) or scrambled shRNA (shGPR137, -) and cultured for 48 h. H The protein expression of GPR137 and hnRPD in A2780 cells transfected with GPR137 shRNA (shGPR137, +) or scrambled shRNA (shGPR137, -). I The protein expression of Flag and hnRPD in OVCAR-8 cells transfected with a vector expressing Flag-GPR137 (Flag-GPR137, +) or a control Flag-tagged empty vector (Flag-GPR137, -). J Western blot analysis for GPR137 and hnRPD in SK-OV-3 cells transfected with GPR137 shRNA (shGPR137, +) or scrambled shRNA (shGPR137, -) and treated for different time periods with CHX. K Western blot analysis for Flag, GPR137 and hnRPD in OVCAR-8 cells transfected with a vector expressing Flag-GPR137 (Flag-GPR137, +) or a control Flag-tagged empty vector (Flag-GPR137, -) and treated for different time periods with CHX. L Western blot analysis for hnRPD in SK-OV-3 cells transfected with GPR137 shRNA (shGPR137, +) or scrambled shRNA (shGPR137, -) and treated with or without MG132. M The mRNA expression of RAB8A in SK-OV-3 cells transfected with hnRPD shRNA or scrambled shRNA (shRNA-Con). ** p < 0.01 versus shRNA-Con; error bar, SD. N = 3. N The mRNA expression of RAB8A in OVCAR-8 cells transfected with a vector expressing hnRPD or a control empty vector (Con). ** p < 0.01 versus Con; error bar, SD. N = 3. O The mRNA levels of RAB8A in SK-OV-3 cells transfected with hnRPD shRNA (sh hnRPD) or scrambled shRNA (shCon) and cultured with actinomycin D (AcD) for the indicated time periods. * p < 0.05; ** p < 0.01 versus shCon; error bar, SD. N = 3. P The mRNA levels of RAB8A in OVCAR-8 cells transfected with a vector expressing hnRPD or a control empty vector (Con) and cultured with actinomycin D (AcD) for the indicated time periods. * p < 0.05; ** p < 0.01 versus Con; error bar, SD. N = 3. Q The mRNA levels of RAB8A in OVCAR-8 cells transfected with a vector expressing GPR137 or a control empty vector (Con) in combination with hnRPD shRNA (sh hnRPD) or scrambled shRNA and cultured with actinomycin D (AcD) for the indicated time periods. * p < 0.05; ** p < 0.01 versus Con; # p < 0.05 versus GPR137; error bar, SD. N = 3. R GEPIA database displayed the correlation between RAB8A and hnRPD

Article Snippet: GPR137 (bs-16270R, 1:1000) antibody and RAB8A (bs-6176R, 1:1000) antibody were acquired from Bioss Biotechnology (Beijing, China), while Flag-tag (M185-3, 1:10000) antibody was obtained from MBL Biotechnology (Beijing, China).

Techniques: Expressing, Co-Immunoprecipitation Assay, Negative Control, Quantitative RT-PCR, Pull Down Assay, Control, Quantitative Proteomics, Transfection, shRNA, Plasmid Preparation, Cell Culture, Western Blot

Journal: Molecular Cancer

Article Title: HnRPD/AUF1 facilitates human ovarian cancer progression through activating FLI1 and maintaining cisplatin resistance

doi: 10.1186/s12943-026-02599-5

Figure Lengend Snippet: GPR137-hnRPD-RAB8A axis regulates OC cell biological behaviors. A CCK-8 assays of OVCAR-8 cells transfected with a GPR137-expressing vector or an empty vector (in Control group) in combination with hnRPD shRNA (shhnRPD) or scrambled shRNA (in Control group) and cultured for the indicated time periods. ** p < 0.01 versus Control, ## p < 0.01 versus GPR137; error bar, SD. N = 3. B Wound healing assays of OVCAR-8 cells transfected with a GPR137-expressing vector or an empty vector (in Control group) in combination with hnRPD shRNA (shhnRPD) or scrambled shRNA (in Control group) at 24 h. Migrated distance was statistically analyzed. Bar, 100 μm. ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. N = 3. C CCK-8 assays of OVCAR-8 cells transfected with a hnRPD-expressing vector or an empty vector (in Control group) in combination with RAB8A shRNA (shRAB8A) or scrambled shRNA (in Control group) and cultured for the indicated time periods. * p < 0.05 versus Control, # p < 0.05 versus hnRPD, ** p < 0.01 versus Control, ## p < 0.01 versus hnRPD; error bar, SD. N = 3. D Wound healing assays of OVCAR-8 cells transfected with a hnRPD-expressing vector or an empty vector (in Control group) in combination with RAB8A shRNA (shRAB8A) or scrambled shRNA (in Control group) at 24 h. Migrated distance was statistically analyzed. Bar, 100 μm. ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+hnRPD; error bar, SD. N = 3. E - H Spearman correlation analysis plots showing the correlation between the pathway scores and the expression of gene hnRPD . In the plots, the x-axis represents the distribution of the expression of hnRPD , and the y-axis represents the distribution of the corresponding pathway score. The values at the top represent the results of the Spearman correlation analysis, including the p -value, and correlation coefficient. I Matrigel invasion assays of OVCAR-8 cells transfected with a GPR137-expressing vector or an empty vector (in Control group) in combination with hnRPD shRNA (shhnRPD) or scrambled shRNA (in Control group) at 24 h. Bar, 100 μm. J Colony formation assays of OVCAR-8 cells infected with lentiviruses expressing GPR137 or control lentiviruses (in Control group) in combination with lentiviruses carrying hnRPD shRNA or scrambled shRNA (in Control group). K Quantitative analysis of (E). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. N = 3. L Quantitative analysis of (F). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. N = 3. M Matrigel invasion assays of OVCAR-8 cells transfected with a hnRPD-expressing vector or an empty vector (in Control group) in combination with RAB8A shRNA (shRAB8A) or scrambled shRNA (in Control group) at 24 h. Bar, 100 μm. N Colony formation assays of OVCAR-8 cells infected with lentiviruses expressing hnRPD or control lentiviruses (in Control group) in combination with lentiviruses carrying RAB8A shRNA or scrambled shRNA (in Control group). O Quantitative analysis of (I). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+hnRPD; error bar, SD. N = 3. P Quantitative analysis of (J). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+hnRPD; error bar, SD. N = 3. Q CCK-8 assays of SK-OV-3 cells transfected with GPR137 shRNA (shGPR137) or scrambled shRNA in combination with a hnRPD-expressing vector or an empty vector (Control) and cultured for the indicated time periods. * p < 0.05 versus Control, # p < 0.05 versus shGPR137; ** p < 0.01 versus Control, ## p < 0.01 versus shGPR137; error bar, SD. R Migrated distance was statistically analyzed in SK-OV-3 cells transfected with GPR137 shRNA (shGPR137) or scrambled shRNA in combination with a hnRPD-expressing vector or an empty vector (Control) at 24 h. ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-GPR137 + Control; error bar, SD. S Matrigel invasion assays of SK-OV-3 cells transfected with GPR137 shRNA (shGPR137) or scrambled shRNA (in Control group) in combination with a hnRPD-expressing vector or an empty vector (in Control group) at 24 h. Bar, 100 μm. T Colony formation assays of SK-OV-3 cells infected with lentiviruses carrying GPR137 shRNA or scrambled shRNA (in Control group) in combination with lentiviruses expressing hnRPD or control lentiviruses (in Control group). U Quantitative analysis of (O). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-GPR137 + Control; error bar, SD. N = 3. V Quantitative analysis of (P). * p < 0.05 versus shRNA-Con+Control, # p < 0.05 versus shRNA-GPR137 + Control; error bar, SD. N = 3

Article Snippet: GPR137 (bs-16270R, 1:1000) antibody and RAB8A (bs-6176R, 1:1000) antibody were acquired from Bioss Biotechnology (Beijing, China), while Flag-tag (M185-3, 1:10000) antibody was obtained from MBL Biotechnology (Beijing, China).

Techniques: CCK-8 Assay, Transfection, Expressing, Plasmid Preparation, Control, shRNA, Cell Culture, Infection

Journal: Molecular Cancer

Article Title: HnRPD/AUF1 facilitates human ovarian cancer progression through activating FLI1 and maintaining cisplatin resistance

doi: 10.1186/s12943-026-02599-5

Figure Lengend Snippet: Suppression of hnRPD expression inhibits OC progression in vivo. A Nude mice with xenografts derived from OVCAR-8 cells stably expressing hnRPD or control. N = 6. B Image showing the size of the tumor xenografts from the two groups in (A). C Weights of xenografts in (B). N = 6. ** p < 0.01 versus Con; error bar, SD. D Tumor volumes of xenografts in (B). N = 6. ** p < 0.01 versus Con; error bar, SD. E The mRNA levels of RAB8A in xenografts in (B). N = 6. ** p < 0.01 versus Con; error bar, SD. F RAB8A staining of RAB8A in xenografts in (B). N = 6. Bar, 20 μm. G IHC score for RAB8A staining in (F). ** p < 0.01 versus Con. H Graphic illustration of the intraperitoneal injection of control or hnRPD stable-overexpressing OVCAR-8 cells. I The metastasis and spread of control or hnRPD stable-overexpressing OVCAR-8 cells were monitored by IVIS Imaging System. N = 5. J Nude mice with xenografts derived from OVCAR-8 cells stably expressing GPR137 or in combination with hnRPD shRNA (shhnRPD). N = 6. K Image showing the size of the tumor xenografts from three groups. L Tumor volume was measured on day 3, 6, 9, 12, 15, and 18, after OVCAR-8 cell injection. N = 6. ** p < 0.01 versus Control, ## p < 0.01 versus GPR137; error bar, SD. M Weights of xenografts derived from OVCAR-8 cells stably expressing GPR137 or in combination with hnRPD shRNA (shhnRPD). N = 6. ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. N Body weight of mice was examined on day 3, 6, 9, 12, 15, and 18, after OVCAR-8 cell injection. ** p < 0.01 versus Control, ## p < 0.01 versus GPR137; error bar, SD. O KI-67 staining and RAB8A staining for tumor tissues derived from OVCAR-8 cells stably expressing GPR137 or in combination with hnRPD shRNA (shhnRPD) on day 18 post inoculation. In Control group, tumor tissues were derived from OVCAR-8 cells infected with control lentiviruses in combination with lentiviruses carrying a scrambled shRNA sequence. Bar, 20 μm. P Statistical analysis of KI-67 index (%) from (F). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. Q IHC score for RAB8A staining in (O). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137. R The mRNA levels of RAB8A in formed tumor tissues derived from OVCAR-8 cells stably expressing GPR137 or in combination with hnRPD shRNA (shhnRPD). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. S The protein levels of RAB8A in formed tumor tissues derived from OVCAR-8 cells stably expressing GPR137 or in combination with hnRPD shRNA (shhnRPD). In Control group, tumor tissues were derived from OVCAR-8 cells infected with control lentiviruses in combination with lentiviruses carrying a scrambled shRNA sequence (labeled as GPR137, -; shhnRPD, -). T qRT-PCR analysis of co-precipitated RAB8A mRNA by hnRPD antibody in the RIP assay in tumor tissues. IgG was used as a negative control. ** p < 0.01 versus IgG; error bar, SD. N = 3. U RNA pull-down assay in tumor tissues with RAB8A RNA or control RNA fragments. Protein abundance was normalized to a Tubulin

Article Snippet: GPR137 (bs-16270R, 1:1000) antibody and RAB8A (bs-6176R, 1:1000) antibody were acquired from Bioss Biotechnology (Beijing, China), while Flag-tag (M185-3, 1:10000) antibody was obtained from MBL Biotechnology (Beijing, China).

Techniques: Expressing, In Vivo, Derivative Assay, Stable Transfection, Control, Staining, Injection, Imaging, shRNA, Infection, Sequencing, Labeling, Quantitative RT-PCR, Negative Control, Pull Down Assay, Quantitative Proteomics