Journal: Orphanet Journal of Rare Diseases

Article Title: Neural stem cells for disease modeling and evaluation of therapeutics for infantile (CLN1/PPT1) and late infantile (CLN2/TPP1) neuronal ceroid lipofuscinoses

doi: 10.1186/s13023-018-0798-2

Figure Lengend Snippet: Effect of recombinant human PPT1/TPP1 protein on reducing enlarged lysosomes in NCL patient NSCs. The western blot analysis ( a , b ) showed that there is a PPT1 deficiency in PPT1 E8/E1 fibroblasts and NSCs, and also there is no TPP1 expression detected in TPP1 E4/E6 and TPP1 E4/IVS5 fibroblast and NSCs. The treatment of NCL NSCs with 200 nM rPPT1/rTPP1 significantly reduced the LysoTracker dye staining( c ), with an effect nearly 99.9% in the NCL NSC lines treated with ERT ( d ). The images were taken with 40X objective lens. Data are displayed as mean ± SD. ** P < 0.01

Article Snippet: Human recombinant PPT1 (14703-H08H) was purchased from Sino Biological (Beijing, China), and TPP1 (2237-SE-010) was ordered from R&D Systems (Minneapolis, MN).

Techniques: Recombinant, Western Blot, Expressing, Staining

Journal: Orphanet Journal of Rare Diseases

Article Title: Neural stem cells for disease modeling and evaluation of therapeutics for infantile (CLN1/PPT1) and late infantile (CLN2/TPP1) neuronal ceroid lipofuscinoses

doi: 10.1186/s13023-018-0798-2

Figure Lengend Snippet: Effect of DT on reducing enlarged lysosomes in NCL patient NSCs. DT dose-dependently reduced the LysoTracker staining ( a ) in NCL NSCs. The quantitative analysis of LysoTracker fluorescence ( b ) revealed that clearance of enlarged lysosomes ranged from 22.8% in the TPP1 E4/E6 NSCs to 33.8% in the PPT1 E8/E1 NSCs after the treatment with 20 μM DT. The images were taken with 40X objective lens. Data are displayed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human recombinant PPT1 (14703-H08H) was purchased from Sino Biological (Beijing, China), and TPP1 (2237-SE-010) was ordered from R&D Systems (Minneapolis, MN).

Techniques: Staining, Fluorescence

Journal: Orphanet Journal of Rare Diseases

Article Title: Neural stem cells for disease modeling and evaluation of therapeutics for infantile (CLN1/PPT1) and late infantile (CLN2/TPP1) neuronal ceroid lipofuscinoses

doi: 10.1186/s13023-018-0798-2

Figure Lengend Snippet: Hydroxypropyl-β-cyclodextrin (HPBCD) ameliorated enlarged lysosomes in NCL NSCs. a Representative image of HPBCD’s effect on reducing LysoTracker staining in NCL NSCs. The maximum reduction of enlarged lysosomes ranged from 31% in TPP1 E4/IVS5 NSCs to 47% in PPT1 E8/E1 NSCs after 1 mM HPBCD treatment ( b ). The images were taken with 40X objective lens. Data are displayed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human recombinant PPT1 (14703-H08H) was purchased from Sino Biological (Beijing, China), and TPP1 (2237-SE-010) was ordered from R&D Systems (Minneapolis, MN).

Techniques: Staining

Journal: Orphanet Journal of Rare Diseases

Article Title: Neural stem cells for disease modeling and evaluation of therapeutics for infantile (CLN1/PPT1) and late infantile (CLN2/TPP1) neuronal ceroid lipofuscinoses

doi: 10.1186/s13023-018-0798-2

Figure Lengend Snippet: The synergistic effect of HPBCD with DT on reduction of enlarged lysosomes in NCL NSCs. a Representative image of a combination of HPBCD and DT on reduction of LysoTracker staining in NCL NSCs compared to DT treatment. b The reduction of enlarged lysosomes ranging from 49% in TPP1 E4/E6 NSCs to 78% in TPP1 E4/IVS5 NSCs. The images were taken with 40X objective lens. Data are displayed as mean ± SD. * P < 0.05, ** P < 0.01

Article Snippet: Human recombinant PPT1 (14703-H08H) was purchased from Sino Biological (Beijing, China), and TPP1 (2237-SE-010) was ordered from R&D Systems (Minneapolis, MN).

Techniques: Staining

Journal: Orphanet Journal of Rare Diseases

Article Title: Neural stem cells for disease modeling and evaluation of therapeutics for infantile (CLN1/PPT1) and late infantile (CLN2/TPP1) neuronal ceroid lipofuscinoses

doi: 10.1186/s13023-018-0798-2

Figure Lengend Snippet: Effect of DT on cytoplasmic lipid droplets accumulation in NCL patient NSCs. DT dose-dependently reduced the Nile red staining ( a ) in NCL NSCs. The quantitative analysis of Nile red fluorescence ( b ) revealed that clearance effect of lipid droplets ranged from 12.4% in the TPP1 E4/E6 NSCs to 34.9% in the TPP1 E4/IVS5 NSCs after the treatment with 20 μM δ-tocopherol. The images were taken with 40X objective lens. Data are displayed as mean ± SD. * P < 0.05, *** P < 0.001 compared to the untreated control

Article Snippet: Human recombinant PPT1 (14703-H08H) was purchased from Sino Biological (Beijing, China), and TPP1 (2237-SE-010) was ordered from R&D Systems (Minneapolis, MN).

Techniques: Staining, Fluorescence, Control

Journal: Orphanet Journal of Rare Diseases

Article Title: Neural stem cells for disease modeling and evaluation of therapeutics for infantile (CLN1/PPT1) and late infantile (CLN2/TPP1) neuronal ceroid lipofuscinoses

doi: 10.1186/s13023-018-0798-2

Figure Lengend Snippet: Co-localization of subunit c and Lamp1 in NCL NSCs and the expression of subunit c in NCL NSCs. a Co-localization of LAMP-1, a lysosomal marker, with subunit c in PPT1 E8/E1 , TPP1 E4/E6 , and wild-type NSCs. The cells were immunostained with antibodies recognizing subunit c (red fluorescence, see white arrows) and Lamp1 (green fluorescence). Minimal overlap of subunit c and Lamp1 immunostaining was observed in wild-type cells (yellow in overlay), but Lamp1 strongly, though not perfectly, overlaps with the accumulated subunit c in PPT1 E8/E1 and TPP1 E4/E6 NSCs. Treatment of INCL and LINCL NSCs with recombinant PPT1 and TPP1 decreased subunit c accumulation in lysosomes of patient cells, respectively. Similar effects were also observed in cells after treatments with δ-tocopherol and HPBCD. Blue represents Hoechst nuclei stain. Images were captured with 60X objective. b and c Expression of subunit c in NCL fibroblasts analyzed by the Western blot. The expressions of subunit c in PPT1 E8/E1 fibroblast were weaker than WT, but the expressions of subunit c increased in TPP1 E4/E6 and TPP1 E4/IVS5 fibroblast compared to WT ( b ). It showed that subunit c expression was decreased by 64% in PPT1 E8/E1 fibroblast, and increased 1.5-fold in both TPP1 E4/E6 and TPP1 E4/IVS5 fibroblast compared to WT ( c ). Data are the mean ± SEM. ** P < 0.01. Expression of subunit c in NCL NSCs (D and E). The expressions of subunit c were weaker in PPT1 E8/E1 NSCs than WT, but the expressions of subunit c were increased in TPP1 E4/E6 and TPP1 E4/IVS5 NSCs compared to WT ( d ). It showed that subunit c expression was decreased by 36% in TPP1 E4/E6 NSCs, and increased 1.2-fold in both TPP1 E4/E6 and TPP1 E4/IVS5 NSCs compared to WT ( e ). Data are displayed as mean ± SD. * P < 0.05, ** P < 0.01, compared to the WT control

Article Snippet: Human recombinant PPT1 (14703-H08H) was purchased from Sino Biological (Beijing, China), and TPP1 (2237-SE-010) was ordered from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Marker, Fluorescence, Immunostaining, Recombinant, Staining, Western Blot, Control

Journal: Orphanet Journal of Rare Diseases

Article Title: Neural stem cells for disease modeling and evaluation of therapeutics for infantile (CLN1/PPT1) and late infantile (CLN2/TPP1) neuronal ceroid lipofuscinoses

doi: 10.1186/s13023-018-0798-2

Figure Lengend Snippet: Effect of DT, HPBCD and enzyme replacement therapy on the accumulation of subunit c in patient NSCs. Cells were treated with 20 μM DT, 1 mM HPBCD, 20 μM DT plus 125 μM HPBCD, or 200 nM rPPT1/rTPP1 for 3 days. PPT1/TPP1 expression were restored after the PPT1/TPP1 replacement therapy. After the treatment with 20 μM DT plus 125 μM HPBCD, the expression of subunit c in PPT1 E8/E1 NSCs decreased by 75% ( a and b ). Moreover, it showed that subunit c expression in TPP1 E4/E6 NSCs and TPP1 E4/IVS5 NSCs was decreased by 51% and 64% respectively, in TPP1 replacement treatment, and also decreased by 18%, 30% with DT treatment ( c , d , e and f ). Data are displayed as mean ± SD. * P < 0.05, ** P < 0.01

Article Snippet: Human recombinant PPT1 (14703-H08H) was purchased from Sino Biological (Beijing, China), and TPP1 (2237-SE-010) was ordered from R&D Systems (Minneapolis, MN).

Techniques: Expressing

Journal: Orphanet Journal of Rare Diseases

Article Title: Neural stem cells for disease modeling and evaluation of therapeutics for infantile (CLN1/PPT1) and late infantile (CLN2/TPP1) neuronal ceroid lipofuscinoses

doi: 10.1186/s13023-018-0798-2

Figure Lengend Snippet: Summary of human cell lines used in the study

Article Snippet: Human recombinant PPT1 (14703-H08H) was purchased from Sino Biological (Beijing, China), and TPP1 (2237-SE-010) was ordered from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Oncology Research

Article Title: Single-Cell and Multi-Omics-Based Characterization of Gastric Cancer Identifies TPP1 as a Potential Target for Gastric Cancer Progression and Treatment

doi: 10.32604/or.2026.070208

Figure Lengend Snippet: This schematic diagram provides an overview of the study’s integrated analytical framework. The workflow begins with data acquisition from public databases, followed by the integration of single-cell RNA sequencing and bulk multi-omics datasets. Metastasis-associated fibroblast subpopulations are then identified through clustering and functional annotation. Key gene modules are extracted using hdWGCNA, and a prognostic risk model is constructed and validated across independent cohorts. Finally, core genes—such as TPP1 —are experimentally validated using in vitro assays to confirm their functional relevance in gastric cancer progression.

Article Snippet: After blocking at room temperature for 1 h, membranes were incubated overnight at 4°C with primary antibodies against TPP1 (1:3000, Proteintech, 25849-1-AP, Chicago, IL, USA) and GAPDH (1:3000, Proteintech, 10494-1-AP, Chicago, IL, USA).

Techniques: Single Cell, RNA Sequencing, Biomarker Discovery, Functional Assay, Construct, In Vitro

Journal: Oncology Research

Article Title: Single-Cell and Multi-Omics-Based Characterization of Gastric Cancer Identifies TPP1 as a Potential Target for Gastric Cancer Progression and Treatment

doi: 10.32604/or.2026.070208

Figure Lengend Snippet: Abnormal expression of TPP1 was verified. ( A ) Boruta algorithm identified six key prognostic signature genes, with TPP1 ranking as the top contributor. ( B ) Pan-cancer differential expression analysis revealed significant overexpression of TPP1 across multiple malignancies. ( C , D ) Bar plots show that TPP1 expression is significantly elevated in gastric cancer tissues based on both unpaired and paired comparisons. ( E ) Kaplan–Meier survival analysis demonstrated that high TPP1 expression correlates with poorer overall survival in gastric cancer patients. ( F ) Immunohistochemical staining confirmed upregulated TPP1 protein expression in gastric cancer tissues (HPA: The Human Protein Atlas). ( G ) Western blot analysis further validated increased TPP1 protein levels in tumor samples. ( H ) qRT-PCR analysis confirmed that TPP1 mRNA expression was significantly elevated in gastric cancer tissues (n = 4). Data are presented as mean ± standard deviation (SD). Statistical comparisons were performed using Student’s t -test unless otherwise indicated. Significance levels were defined as follows: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: After blocking at room temperature for 1 h, membranes were incubated overnight at 4°C with primary antibodies against TPP1 (1:3000, Proteintech, 25849-1-AP, Chicago, IL, USA) and GAPDH (1:3000, Proteintech, 10494-1-AP, Chicago, IL, USA).

Techniques: Expressing, Quantitative Proteomics, Over Expression, Immunohistochemical staining, Staining, Western Blot, Quantitative RT-PCR, Standard Deviation

Journal: Oncology Research

Article Title: Single-Cell and Multi-Omics-Based Characterization of Gastric Cancer Identifies TPP1 as a Potential Target for Gastric Cancer Progression and Treatment

doi: 10.32604/or.2026.070208

Figure Lengend Snippet: Silencing of TPP1 attenuates tumorigenic properties and enhances apoptosis in gastric cancer cells. ( A ) qRT-PCR analysis confirmed elevated TPP1 expression in gastric cancer cell lines (n = 3). ( B ) Western blot analysis validated high TPP1 protein levels across gastric cancer cell lines (n = 3). ( C ) qRT-PCR confirmed effective knockdown of TPP1 following siRNA transfection (n = 3). ( D ) Colony formation assays demonstrated a significant reduction in clonogenic capacity upon TPP1 silencing (n = 3). ( E ) Flow cytometry analysis using Annexin V/DAPI staining revealed increased apoptosis in TPP1-depleted cells (n = 3). ( F ) Transwell invasion assays showed a marked decrease in invasive capacity following TPP1 knockdown (n = 3). ( G ) CCK-8 assays indicated impaired cell viability in TPP1-silenced gastric cancer cells (n = 3). Data are presented as mean ± standard deviation (SD), and all experiments were performed in triplicate. Statistical significance was assessed using Student’s t-test unless otherwise specified: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).

Article Snippet: After blocking at room temperature for 1 h, membranes were incubated overnight at 4°C with primary antibodies against TPP1 (1:3000, Proteintech, 25849-1-AP, Chicago, IL, USA) and GAPDH (1:3000, Proteintech, 10494-1-AP, Chicago, IL, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Knockdown, Transfection, Flow Cytometry, Staining, CCK-8 Assay, Standard Deviation