Journal: PLoS ONE

Article Title: Transmigration of Neural Stem Cells across the Blood Brain Barrier Induced by Glioma Cells

doi: 10.1371/journal.pone.0060655

Figure Lengend Snippet: A) Transmigration of ReNcells CX across RBMECs induced by astrocyte or glioma C6 CM. Light microscopy image of toluidine blue stained cells on the basal surface of a filter and scheme illustrating each assay. B) Graphed data. N = 3, F(1,12) = 343.8; *** p<0.001, ns = not significant; as assessed by two-way ANOVA followed by Bonferroni's post hoc test. C) In an 8 h transmigration assay, a similar degree of ReNcells CX transmigration is attained with glioma C6 cells or glioma C6 CM placed in the basal compartment. N = 10, F(3,36) = 60.74; ***P<0.001, ns = not significant; as assessed by one-way ANOVA followed by Bonferroni's post hoc test. D) Representative light microscopy image of toluidine blue stained cells on the basal surface of the filter and scheme illustrating each assay.

Article Snippet: Quantification was done in 4 day CM derived from astrocytes and glioma C6 cells using solid phase sandwich ELISA for VEGF (Invitrogen, KHG0111), HGF (Ray Biotech Inc., ELM-HGF-001), EGF (Peprotech, 900-M390) and PGE 2 (ENZO Life Science, Farmingdale, N.Y., ADI-900-001).

Techniques: Transmigration Assay, Light Microscopy, Staining

Journal: PLoS ONE

Article Title: Transmigration of Neural Stem Cells across the Blood Brain Barrier Induced by Glioma Cells

doi: 10.1371/journal.pone.0060655

Figure Lengend Snippet: A) TEER of RBMECs in the conditions illustrated in the scheme. N = 3, F(3,30) = 19.97; *P<0.05, **P<0.01, ***P<0.001, with respect to 1; # P<0.05, ## P<0.01, ### P<0.001, with respect to 3; as assessed by two-way ANOVA followed by Bonferroni's post hoc test. B) Immunofluorescence localization of claudin-5 and occludin in RBMECs incubated with astrocytes or glioma C6 CM. Arrow, continuous cell border staining; arrowheads, holes appearing at cell borders. C) Scanning electron micrograph of ReNcells CX moving across a hole in the endothelial monolayer.

Article Snippet: Quantification was done in 4 day CM derived from astrocytes and glioma C6 cells using solid phase sandwich ELISA for VEGF (Invitrogen, KHG0111), HGF (Ray Biotech Inc., ELM-HGF-001), EGF (Peprotech, 900-M390) and PGE 2 (ENZO Life Science, Farmingdale, N.Y., ADI-900-001).

Techniques: Immunofluorescence, Incubation, Staining

Journal: PLoS ONE

Article Title: Transmigration of Neural Stem Cells across the Blood Brain Barrier Induced by Glioma Cells

doi: 10.1371/journal.pone.0060655

Figure Lengend Snippet: A) The amount of HGF and VEGF determined by ELISA is higher in glioma C6 CM than in astrocyte CM, while the opposite is found for EGF. IP of HGF, VEGF or EGF reduces the amount of the growth factor present in CM. HGF: N = 4, F(3,12) = 26.54; VEGF: N = 3, F(3,8) = 20.98; EGF: N = 3, F(3,8) = 44.52; **P<0.01, ***P<0.001, ns = not significant; as assessed by one-way ANOVA followed by Bonferroni's post hoc test. B) ReNcells CX transmigration assay across RBMECs done by placing in the basal compartment of the Millicell, DMEM with HGF or EGF; or DMEM with neutralizing antibodies against HGF, VEGF or EGF; or astrocyte or glioma C6 CM with or without neutralizing antibodies against HGF, VEGF and EGF. Control corresponds to the transmigration induced by glioma C6 CM without the neutralizing antibodies against HGF (upper panel), VEGF (middle panel) and EGF (lower panel), and is considered as 100% transmigration. HGF: N = 3, F(6,14) = 124.7; VEGF: N = 3, F(4,10) = 275.5; EGF: N = 3, F(7,16) = 30.77; **P<0.01, ***P<0.001, ns = not significant; as assessed by one-way ANOVA followed by Bonferroni's post hoc test. C) RBMECs cultured for 8 h in DMEM with HGF and EGF have a TEER similar to that obtained with astrocyte CM. IP of VEGF reverses the decrease in TEER induced by glioma C6 CM whereas IP of EGF from astrocyte CM decreases TEER in a manner similar to that obtained with glioma C6 CM. HGF: N = 3, F(2,16) = 5.668; **P<0.01 with respect to astrocytes CM; VEGF: N = 3, F(2,20) = 14.64; *P<0.05 with respect to astrocytes CM; #P<0.05 with respect to glioma C6 CM; EGF: N = 3, F(3,30) = 23.07; *P<0.05, **P<0.01, ***P<0.001 with respect to astrocytes CM; # P<0.05, ## P<0.01, ### P<0.001 with respect to glioma C6 CM; + P<0.05, ++ P<0.01, +++ P<0.001 with respect to astrocytes CM + α-EGF; as assessed by one-way ANOVA followed by Bonferroni's post hoc test.

Article Snippet: Quantification was done in 4 day CM derived from astrocytes and glioma C6 cells using solid phase sandwich ELISA for VEGF (Invitrogen, KHG0111), HGF (Ray Biotech Inc., ELM-HGF-001), EGF (Peprotech, 900-M390) and PGE 2 (ENZO Life Science, Farmingdale, N.Y., ADI-900-001).

Techniques: Enzyme-linked Immunosorbent Assay, Transmigration Assay, Control, Cell Culture

Journal: PLoS ONE

Article Title: Transmigration of Neural Stem Cells across the Blood Brain Barrier Induced by Glioma Cells

doi: 10.1371/journal.pone.0060655

Figure Lengend Snippet: A) Quantitation of PGE 2 by ELISA in astrocyte CM and in glioma C6 CM derived from cells treated or not with COX-2 inhibitor NS398. N = 3, F(2,6) = 161.9; **P<0.01, ***P<0.001; as assessed by one-way ANOVA followed by Bonferroni's post hoc test. B) TEER of RBMECs cultures incubated in the basal compartment with astrocytes or glioma C6 CM derived from control or NS398 treated cells. N = 3, F(2,20) = 53.56; *P<0.05, **P<0.01 with respect to astrocytes CM; ## P<0.01, ### P<0.001 with respect to glioma C6 CM; as assessed by two-way ANOVA followed by Bonferroni's post hoc test. C) Quantitative analysis of cytokines IFN-α, TNF-α, IL-12p70, IL-1β, IL-6, IL-8 and IL-10 present in astrocyte and glioma C6 CM. N = 3, df = 2, as assessed by Student's t-test.

Article Snippet: Quantification was done in 4 day CM derived from astrocytes and glioma C6 cells using solid phase sandwich ELISA for VEGF (Invitrogen, KHG0111), HGF (Ray Biotech Inc., ELM-HGF-001), EGF (Peprotech, 900-M390) and PGE 2 (ENZO Life Science, Farmingdale, N.Y., ADI-900-001).

Techniques: Quantitation Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Incubation, Control

Journal: PLoS ONE

Article Title: Transmigration of Neural Stem Cells across the Blood Brain Barrier Induced by Glioma Cells

doi: 10.1371/journal.pone.0060655

Figure Lengend Snippet: A) Zonulin is detected by Western blot in glioma C6 CM and not in astrocyte CM. B) ReNcells CX transmigration assay across RBMECs done by placing in the basal compartment of the Millicell, astrocyte or glioma C6 CM with or without zonulin. N = 6, F(3,20) = 228.2; *** p<0.001; as assessed by one-way ANOVA followed by Bonferroni's post hoc test. C) Elimination of zonulin from glioma C6 CM reverses the decrease in TEER exerted by glioma C6 CM. N = 3, F(3,30) = 5.981; **P<0.01, ***P<0.001 with respect to glioma C6 CM; as assessed by two-way ANOVA followed by Bonferroni's post hoc test.

Article Snippet: Quantification was done in 4 day CM derived from astrocytes and glioma C6 cells using solid phase sandwich ELISA for VEGF (Invitrogen, KHG0111), HGF (Ray Biotech Inc., ELM-HGF-001), EGF (Peprotech, 900-M390) and PGE 2 (ENZO Life Science, Farmingdale, N.Y., ADI-900-001).

Techniques: Western Blot, Transmigration Assay

Journal: Frontiers in Oncology

Article Title: Spatial transcriptome and single-cell sequencing reveal the role of nucleotide metabolism in breast cancer progression and tumor microenvironment

doi: 10.3389/fonc.2025.1703778

Figure Lengend Snippet: Predicting therapy response with NMRS and validation of the core genes. (A) Estimated IC50 values of Tamoxifen and Palbociclib in high- versus low NMRS groups. (B) Estimated IC50 values of Olaparib and Gefitinib in high- versus low NMRS groups. (C, D) The correlation and differential analysis of drug sensitivity for potential drugs screened from the CTRP and PRISM datasets. (E) Validation of the expression of TAGLN2, PCMT1, PTMA, TUBA3D, and DCTPP1 in a normal breast cell line (MCF-10A) and three BC cell lines (MDA-MB-231, MCF7, and SK-BR3) by qRT-PCR. (F) Immunohistochemical images showing the protein expression of the five genes in the HPA database. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05, ns p > 0.05. BC breast cancer, HPA Human Protein Atlas.

Article Snippet: The antibodies used in this study include: DCTPP1 (Proteintech, # 16684-1-AP, 1:1000), β-Actin (Proteintech, #66009-1-Ig, 1:5000) and anti-Rabbit secondary antibody (Proteintech, Cat: #SA00001-2, 1:5000).

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Immunohistochemical staining

Journal: Frontiers in Oncology

Article Title: Spatial transcriptome and single-cell sequencing reveal the role of nucleotide metabolism in breast cancer progression and tumor microenvironment

doi: 10.3389/fonc.2025.1703778

Figure Lengend Snippet: DCTPP1 expression and functional validation in BC (A) DCTPP1 mRNA expression was higher in TCGA-BRCA tumor tissues than in normal tissues ( p < 0.01). (B) Overall survival was worse in TCGA-BRCA patients with high DCTPP1 expression ( p < 0.0001). (C) ROC curve analysis of DCTPP1, highlighting its diagnostic potential. (D) Single-cell analysis showed ( GSE161529 ) that DCTPP1 expression was significantly higher in NUhighepi cells than in others. (E, F) WB and qpcr analysis of the knockdown efficiency of DCTPP1 in MDA-MB-231 (E) and SK-BR-3 (F) cells and showed that the DCTPP1 gene was successfully knocked down in MDA-MB-231 and SK-BR-3 cell lines. (G) CCK-8 showed that the proliferation activity of the cells that knockdown DCTPP1 was dramatically reduced. (H) After DCTPP1 knockdown, the cloning ability of MDA-MB-231 and SK-BR-3 cell lines decreased significantly. (I) Healing test. After DCTPP1 knockdown, the migration ability of MDA-MB-231 and SK-BR-3 cell lines decreased significantly. (J) Transwell assay. After DCTPP1 knockdown, the migration and invasion abilities of MDA-MB-231 and SK-BR-3 cell lines were significantly decreased. **** p < 0.0001, *** p < 0.001, * p < 0.05, ns p > 0.05.

Article Snippet: The antibodies used in this study include: DCTPP1 (Proteintech, # 16684-1-AP, 1:1000), β-Actin (Proteintech, #66009-1-Ig, 1:5000) and anti-Rabbit secondary antibody (Proteintech, Cat: #SA00001-2, 1:5000).

Techniques: Expressing, Functional Assay, Biomarker Discovery, Diagnostic Assay, Single-cell Analysis, Knockdown, CCK-8 Assay, Activity Assay, Cloning, Migration, Transwell Assay