Journal: Cancers

Article Title: Protein Disulphide Isomerase A1 Is Involved in the Regulation of Breast Cancer Cell Adhesion and Transmigration via Lung Microvascular Endothelial Cells

doi: 10.3390/cancers12102850

Figure Lengend Snippet: Effects of PDIA1 inhibition on wound-healing and migration of breast cancer cells and endothelial cells. ECIS Wound-Healing Assay of hLMVEC, MCF-7 and MDA-MB-231 cells and breast cancer sublines with silencing of PDIA1 (shN, shPDIA1-1 and shPDIA1-3). Real time tracings in hLMVECs ( A ), MCF-7 ( C ) and MDA-MB-231 ( E ) cell lines after addition of bepristat 2a at various concentrations (1, 10, 30 or 50 µM) and after PDIA1 silencing in MCF-7 ( G ) and MDA-MB-231 ( I ) cell lines. Area under the curve boxplots represent AUC quantitation of changes in migration rate of bepristat 2a-treated hLMVECs ( B ), MCF-7 ( D ) and MDA-MB-231 ( F ) cell lines versus non-treated controls as well as MCF-7 ( H ) or MDA-MB-231 ( J ) sublines transduced against PDIA1 (shPDIA1-1, shPDIA1-3) or wild type cells regarding to negative sequence (shN). The line graphs and AUC boxplots represent mean ± SD of three independent experiments. Statistical analysis was calculated using parametric one-way ANOVA followed by Dunnett’s multiple comparisons test (* p = 0.05, ** p = 0.01, *** p = 0.001).

Article Snippet: Human lung microvascular endothelial cell line (hLMVECs) was obtained from Cell Applications (San Diego, CA, USA).

Techniques: Inhibition, Migration, Wound Healing Assay, Quantitation Assay, Sequencing

Journal: Cancers

Article Title: Protein Disulphide Isomerase A1 Is Involved in the Regulation of Breast Cancer Cell Adhesion and Transmigration via Lung Microvascular Endothelial Cells

doi: 10.3390/cancers12102850

Figure Lengend Snippet: Effects of exogenous PDIA1 and PDIA3 on adhesive interaction between breast cancer cells and different substrates. Effect of exogenous proteins PDIA1 and PDIA3 on adhesion of MCF-7 ( A – F ) and MDA-MB-231 ( G – L ) cells to collagen type I, fibronectin and lung microvascular hLMVEC cells, respectively. Data represent mean ± SD of three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s multiple comparisons test (* p = 0.05, ** p = 0.01, *** p = 0.001).

Article Snippet: Human lung microvascular endothelial cell line (hLMVECs) was obtained from Cell Applications (San Diego, CA, USA).

Techniques: Adhesive

Journal: Cellular and molecular bioengineering

Article Title: Neurogenic potential of engineered mesenchymal stem cells overexpressing VEGF

doi: 10.1007/s12195-015-0425-4

Figure Lengend Snippet: Endogenous VEGF protein expression was upregulated 4 days post-sciatic nerve transection. Squares, sham surgery. Circles, nerve transection. Data are mean ± SEM, n=4, *p<0.01.

Article Snippet: As a positive control, DRGs were cultured in basal media supplemented with 5 or 100 ng/mL human recombinant VEGF (R&D systems).

Techniques: Expressing

Journal: Cellular and molecular bioengineering

Article Title: Neurogenic potential of engineered mesenchymal stem cells overexpressing VEGF

doi: 10.1007/s12195-015-0425-4

Figure Lengend Snippet: Co-culture of DRG explants, SCs and HAECs with control or VEGF-MSCs increased neurite outgrowth and cell proliferation. (A) Neurite length was increased in DRG explants in co-cultures with control and VEGF-MSCs (*p<0.01 versus basal media). (B) SC proliferation was increased in co-cultures with control and VEGF-MSCs (*p<0.01 versus basal media). (C) HAEC proliferation was increased in co-cultures with control and VEGF-MSCs compared to basal media, and co-cultures with VEGF-MSCs increased proliferation compared to control MSCs (*p<0.01, n≥6). (D–F) Supplementation of cultures with human recombinant VEGF increased cell proliferation in (F) HAEC cultures (*p<0.01) but not (D) DRG explants or (E) SC cultures (n≥7). Data are normalized to their respective basal media controls. Data are mean ± SEM.

Article Snippet: As a positive control, DRGs were cultured in basal media supplemented with 5 or 100 ng/mL human recombinant VEGF (R&D systems).

Techniques: Co-Culture Assay, Control, Recombinant

Journal: Cellular and molecular bioengineering

Article Title: Neurogenic potential of engineered mesenchymal stem cells overexpressing VEGF

doi: 10.1007/s12195-015-0425-4

Figure Lengend Snippet: Addition of the VEGF receptor inhibitor, SU5416, reduced proliferation of HAECs but did not effect neurite extension or SC proliferation. (A) Neurite length of DRG explant cultures. (B) Quantification of SC proliferation. (C) Quantification of HAEC proliferation. Filled columns represent data in the absence of SU5416; open columns represent data in the presence of 0.5 μM SU5416. Data are normalized to their respective basal media controls. Data are mean ± SEM, n≥6, *p<0.01.

Article Snippet: As a positive control, DRGs were cultured in basal media supplemented with 5 or 100 ng/mL human recombinant VEGF (R&D systems).

Techniques:

Journal: Cellular and molecular bioengineering

Article Title: Neurogenic potential of engineered mesenchymal stem cells overexpressing VEGF

doi: 10.1007/s12195-015-0425-4

Figure Lengend Snippet: VEGF-MSCs maintained their overexpression of growth factor and proliferation rate in 3D fibrin gels. (A) VEGF-MSCs maintain VEGF overexpression on tissue culture plastic (TCP) and in fibrin gels. Filled columns: control MSCs; open columns: VEGF-MSCs (n=6, *p<0.01 versus control MSCs). (B) Proliferation of control and VEGF-MSCs was not different when cultured on TCP or in fibrin gels. Filled columns, control MSCs. Open columns, VEGF-MSCs. (n=4). (C) Conduits filled with VEGF-MSCs embedded within fibrin gels secrete detectable levels of VEGF for up to 21 days (n=4). Data are mean ± SEM. Fluorescent images of conduits filled with green fluorescent protein (GFP)-positive VEGF-MSCs embedded within fibrin gels at Day (D) 0, (E) 6, (F) 12, and (G) 21. Scale bar=100 μm.

Article Snippet: As a positive control, DRGs were cultured in basal media supplemented with 5 or 100 ng/mL human recombinant VEGF (R&D systems).

Techniques: Over Expression, Control, Cell Culture

Journal: Cellular and molecular bioengineering

Article Title: Neurogenic potential of engineered mesenchymal stem cells overexpressing VEGF

doi: 10.1007/s12195-015-0425-4

Figure Lengend Snippet: GFP-positive MSCs remained localized at the site of implantation at week 2. Confocal images of transected sciatic treated with a PLLA conduit containing (A) fibrin alone, (B) a fibrin matrix with control MSCs, or (C) a fibrin matrix with VEGF-MSCs. Conduits are oriented with the proximal end on the left and the distal end on the right. Green, GFP for endogenous green fluorescent protein positive-MSCs. Blue, DAPI for cell nuclei. Scale bar = 500 μm.

Article Snippet: As a positive control, DRGs were cultured in basal media supplemented with 5 or 100 ng/mL human recombinant VEGF (R&D systems).

Techniques: Control

Journal: Cellular and molecular bioengineering

Article Title: Neurogenic potential of engineered mesenchymal stem cells overexpressing VEGF

doi: 10.1007/s12195-015-0425-4

Figure Lengend Snippet: Conduits filled with VEGF-MSCs embedded within fibrin gels maintain VEGF overexpression 2 weeks post-transplantation. Filled columns: control MSCs; open columns: VEGF-MSCs. Data are mean ± SEM, n≥3, *p<0.01.

Article Snippet: As a positive control, DRGs were cultured in basal media supplemented with 5 or 100 ng/mL human recombinant VEGF (R&D systems).

Techniques: Over Expression, Transplantation Assay, Control

Journal: Cell reports

Article Title: Two-pore channel-2 and inositol trisphosphate receptors coordinate Ca 2+ signals between lysosomes and the endoplasmic reticulum

doi: 10.1016/j.celrep.2023.113628

Figure Lengend Snippet: (A‒C) Effect of TPC2-A1-N or TPC2-A1-P on histamine-induced Ca 2+ signals in HeLa cells loaded with Fura-2. (A) Time course showing the effect of co-stimulating cell populations in an automated manner with increasing concentrations of histamine and a fixed concentration of DMSO (0.1% [v/v]), TPC2-A1-N (N, 30 μM), or TPC2-A1-P (P, 60 μM). (B) Concentration-effect relationship for peak histamine responses in the presence of indicated reagents. (C) Effect of co-stimulating cells with increasing concentrations of TPC2-A1-N or TPC2-A1-P at a fixed concentration of histamine (0.8 μM, left, or 2.6 μM, right). Data are expressed as mean ± SEM from 3–4 biological replicates. (D and E) Effect of TPC2-A1-N or TPC2-A1-P on histamine-induced Ca 2+ signals in U2OS cells loaded with Fura-2. (D) Time course showing the effect of co-stimulating cell populations in an automated manner with increasing concentrations of histamine and a fixed concentration of DMSO (0.1% [v/v]) or TPC2-A1-N (10 μM). (E) Concentration-effect relationship for peak histamine responses in the presence of TPC2-A1-N (10 μM) or TPC2-A1-P (20 μM). Data are expressed as mean ± SEM from 3–4 biological replicates. (F and G) Effect of TPC2-A1-N on histamine-induced NO production in rat brain microvascular endothelial cells (RBMVECs) loaded with DAF 2A. (F) Time course data showing the effect of co-stimulating cells with increasing concentrations of histamine and a fixed concentration of DMSO (0.1% [v/v]) or TPC2-A1-N (N, 10 μM). Each trace is the fluorescence response of a single cell. The thicker trace is the average of the population. (G) Concentration-effect relationship for peak histamine responses in the presence of indicated reagents. Data were corrected for basal NO production in the absence of histamine. Data are expressed as mean ± SEM from 27–33 cells. ****p < 0.0001, n.s., not significant, two-way ANOVA followed by Bonferroni’s test. (H) Proposed model whereby local Ca 2+ signals stemming from the lysosome via TPC2 and the ER via IP 3 receptors form regional intermediaries that precede global Ca 2+ signals upon cellular stimulation. Created with BioRender.com .

Article Snippet: Rat brain microvascular endothelial cells (Cell Applications, Inc. (San Diego, CA, USA) were cultured in rat brain endothelial cell basal medium and rat brain endothelial cell growth supplement in flasks coated with attachment factor according to the manufacturer’s instructions (Cell Applications, Inc.) , For single cell epifluorescence imaging, HeLa and U2OS cells were plated onto round 13 mm diameter coverslips (Academy) coated with poly-L-lysine (20 μg/mL, Sigma).

Techniques: Concentration Assay, Fluorescence, Cell Stimulation

Journal: Cell reports

Article Title: Two-pore channel-2 and inositol trisphosphate receptors coordinate Ca 2+ signals between lysosomes and the endoplasmic reticulum

doi: 10.1016/j.celrep.2023.113628

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rat brain microvascular endothelial cells (Cell Applications, Inc. (San Diego, CA, USA) were cultured in rat brain endothelial cell basal medium and rat brain endothelial cell growth supplement in flasks coated with attachment factor according to the manufacturer’s instructions (Cell Applications, Inc.) , For single cell epifluorescence imaging, HeLa and U2OS cells were plated onto round 13 mm diameter coverslips (Academy) coated with poly-L-lysine (20 μg/mL, Sigma).

Techniques: Recombinant, CRISPR, Plasmid Preparation, Software