Journal: Journal of Biological Chemistry

Article Title: Evidence That Factor VIII Forms a Bivalent Complex with the Low Density Lipoprotein (LDL) Receptor-related Protein 1 (LRP1)

doi: 10.1074/jbc.m116.754622

Figure Lengend Snippet: FIGURE 7. Effect of clusters II and IV on LRP1-mediated internalization of BDD-fVIII (A) and tPAPAI-1 (B) by WI-38 fibroblasts. WI-38 fibroblasts were seeded in 12-well tissue culture dishes (1 105 cells/well) and incubated with 20 nM 125I-BDD-fVIII (A) or 10 nM 125I-tPAPAI-1 (B) for 4 h at 37 °C in the presence orabsenceofLRP1clusterIIorIV(500nM).Followingincubation,theamountofradiolabeledligandinternalizedwasdetermined,andthedatawerenormalized to the amount of fVIII internalized in the absence of competitors. The results represent the mean of three independent experiments. Error bars represent S.E. (*, p 0.002, one-way analysis of variance with Tukey’s multiple comparison post-test).

Article Snippet: WI38 cells, human lung fibroblast cells, were obtained from American Type Culture Collection (ATCC) and maintained in DMEM (Corning) supplemented with 10% FBS and penicillin/streptomycin.

Techniques: Incubation, Comparison

Journal: The Journal of Clinical Investigation

Article Title: SIRT2 protects peripheral neurons from cisplatin-induced injury by enhancing nucleotide excision repair

doi: 10.1172/JCI123159

Figure Lengend Snippet: (A) Sirt2 KO in primary DRG neurons resulted in decreased cell survival after cisplatin treatment. Data were analyzed by 2-tailed Student’s t test (n = 3). (B and C) The viability of neuronally differentiated 50B11 (B) and PC12 (C) cells after various doses of cisplatin treatment was measured with trypan blue staining. Reexpression of WT-SIRT2, but not the enzymatically inactive mutant, HY-SIRT2, in Sirt2-KO 50B11 and PC12 cells increased cell resistance to cisplatin cytotoxicity. One-way ANOVA demonstrated a main effect for cell genotype (P < 0.001). *P < 0.05; **P < 0.01; ***P < 0.001 denote significance levels detected among various cell genotypes and at different time points by Tukey’s post hoc analysis. (D and E) The viability of control vector and Sirt2-KO LLC (D) and H1299 (E) cells after varying doses of cisplatin treatment, as measured by trypan blue staining. Sirt2 KO in LLC and H1299 cells shows no difference in cell viability following cisplatin treatment, as analyzed by 2-tailed Student’s t test.

Article Snippet: The mouse LLC cell line LL/2 (LLC1), human non–small cell lung cancer cell line H1299, and rat pheochromocytoma cell line PC12 were purchased from ATCC.

Techniques: Staining, Mutagenesis, Plasmid Preparation

Journal: Oncology Letters

Article Title: Polyphyllin I and VII potentiate the chemosensitivity of A549/DDP cells to cisplatin by enhancing apoptosis, reversing EMT and suppressing the CIP2A/AKT/mTOR signaling axis

doi: 10.3892/ol.2019.10895

Figure Lengend Snippet: Characterization of A549 and A549/DDP cells. (A) A549 and A549/DDP cells were exposed to different concentrations of DDP (1–32 µg/ml) for 24 h, and cell viability was determined by Cell Counting Kit-8 assay. The protein expression levels of (B) E-cadherin, vimentin, α-SMA and (C) CIP2A, p-AKT, AKT, p-mTOR were measured using western blotting; E, epithelial; SMA, smooth muscle actin; p-mTOR, phosphorylated mammalian target of rapamycin; AKT, protein kinase B; DDP, cisplatin; CIP2A, cancerous inhibitor of protein phosphatase 2A; IC 50 , half maximal inhibitory concentration.

Article Snippet: Human NSCLC cell lines A549 and DDP-resistant A549/DDP cells (OriGene Technologies, Inc.) were cultured in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and penicillin (100 U/ml)-streptomycin (100 mg/ml) at 37°C, with 5% CO 2 .

Techniques: Cell Counting, Expressing, Western Blot, Concentration Assay

Journal: Oncology Letters

Article Title: Polyphyllin I and VII potentiate the chemosensitivity of A549/DDP cells to cisplatin by enhancing apoptosis, reversing EMT and suppressing the CIP2A/AKT/mTOR signaling axis

doi: 10.3892/ol.2019.10895

Figure Lengend Snippet: PPI and PPVII induce cytotoxicity in A549 and A549/DDP cells. (A) Chemical structure of PPI and PPVII. (B) A549 and A549/DDP cells were treated with different dose of PPI (0.5–8 µg/ml) and PPVII (0.5–8 µg/ml) for 24 h, respectively. In A549 cells, the cell inhibition rate of 0.5 µg/ml PPI was significantly different from other concentration groups (**P<0.01); 1.0 µg/ml was significantly different from 4.0 and 8.0 µg/ml (**P<0.01), and 2.0 µg/ml was significantly different from 8.0 µg/ml (**P<0.01). As for A549/DDP cells, the cell inhibition rates of 0.5 µg/ml PPI and 1.0 µg/ml were both significantly different from other concentration groups ( ## P<0.01), and 2.0 µg/ml was significantly different from 8.0 µg/ml ( ## P<0.01). In A549 cells, there were significant differences in cell inhibition rates of PPVII among different concentration groups (**P<0.01); as for A549/DDP cells, there were significant differences among different concentration groups except for 4.0 and 8.0 µg/ml ( ## P<0.01). PPI, Polyphyllin I; PPVII, polyphyllin VII; DDP, cisplatin.

Article Snippet: Human NSCLC cell lines A549 and DDP-resistant A549/DDP cells (OriGene Technologies, Inc.) were cultured in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and penicillin (100 U/ml)-streptomycin (100 mg/ml) at 37°C, with 5% CO 2 .

Techniques: Inhibition, Concentration Assay

Journal: Oncology Letters

Article Title: Polyphyllin I and VII potentiate the chemosensitivity of A549/DDP cells to cisplatin by enhancing apoptosis, reversing EMT and suppressing the CIP2A/AKT/mTOR signaling axis

doi: 10.3892/ol.2019.10895

Figure Lengend Snippet: PPI and PPVII possess chemo-sensitizing effects on A549/DDP cells. (A) A549/DDP cells treated with indicated doses of DDP, or DDP plus either 0.375 µg/ml PPI or 0.75 µg/ml PPI, respectively for 24 h. (B) A549/DDP cells treated with indicated doses of DDP, or DDP plus either 0.375 µg/ml PPVII or 0.75 µg/ml PPVII, respectively for 24 h, cell viability was measured with Cell Counting Kit-8 assay. Data were presented as the mean ± standard deviation. *P<0.05, **P<0.01 and ***P<0.001 vs. DDP treated group. # P<0.05 and ## P<0.01. PPI, Polyphyllin I; PPVII, polyphyllin VII; DDP, cisplatin.

Article Snippet: Human NSCLC cell lines A549 and DDP-resistant A549/DDP cells (OriGene Technologies, Inc.) were cultured in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and penicillin (100 U/ml)-streptomycin (100 mg/ml) at 37°C, with 5% CO 2 .

Techniques: Cell Counting, Standard Deviation

Journal: Oncology Letters

Article Title: Polyphyllin I and VII potentiate the chemosensitivity of A549/DDP cells to cisplatin by enhancing apoptosis, reversing EMT and suppressing the CIP2A/AKT/mTOR signaling axis

doi: 10.3892/ol.2019.10895

Figure Lengend Snippet: PPI and PPVII reduce the IC 50 value of cisplatin in A549/DDP cells

Article Snippet: Human NSCLC cell lines A549 and DDP-resistant A549/DDP cells (OriGene Technologies, Inc.) were cultured in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and penicillin (100 U/ml)-streptomycin (100 mg/ml) at 37°C, with 5% CO 2 .

Techniques:

Journal: Oncology Letters

Article Title: Polyphyllin I and VII potentiate the chemosensitivity of A549/DDP cells to cisplatin by enhancing apoptosis, reversing EMT and suppressing the CIP2A/AKT/mTOR signaling axis

doi: 10.3892/ol.2019.10895

Figure Lengend Snippet: PPI and PPVII enhance DDP-induced apoptosis. A549/DDP cells were treated with DDP (6 µg/ml), PPI (0.75 µg/ml), PPVII (0.75 µg/ml), or DDP (6 µg/ml) in combination with either PPI (0.75 µg/ml) or PPVII (0.75 µg/ml) for 24 h, respectively. Apoptosis of cells was detected by Annexin V-FITC/PI staining and flow cytometry. The apoptotic percentage is shown as a bar graph. Data represents the mean ± standard deviation of three independent experiments. **P<0.01 and ***P<0.001. PPI, Polyphyllin I; PPVII, polyphyllin VII; DDP, cisplatin; FITC, fluorescein isothiocyanate; PI, propidium iodide.

Article Snippet: Human NSCLC cell lines A549 and DDP-resistant A549/DDP cells (OriGene Technologies, Inc.) were cultured in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and penicillin (100 U/ml)-streptomycin (100 mg/ml) at 37°C, with 5% CO 2 .

Techniques: Staining, Flow Cytometry, Standard Deviation

Journal: Oncology Letters

Article Title: Polyphyllin I and VII potentiate the chemosensitivity of A549/DDP cells to cisplatin by enhancing apoptosis, reversing EMT and suppressing the CIP2A/AKT/mTOR signaling axis

doi: 10.3892/ol.2019.10895

Figure Lengend Snippet: PPI and PPVII enhance DDP-induced apoptosis through the P53 pathway and caspases-dependent pathway. A549/DDP cells were treated with DDP (6 µg/ml), PPI (0.75 µg/ml), PPVII (0.75 µg/ml), or DDP (6 µg/ml) in combination with either PPI (0.75 µg/ml) or PPVII (0.75 µg/ml) for 24 h, respectively. The protein expression levels of P53, Bax, Bcl-2 (A), PARP, C-PARP, pro-Caspase-3, C-Caspase-3 (B) were measured using western blotting. PPI, Polyphyllin I; PPVII, polyphyllin VII; DDP, cisplatin; C-PARP, cleaved-poly (ADP-ribose) polymerase 1.

Article Snippet: Human NSCLC cell lines A549 and DDP-resistant A549/DDP cells (OriGene Technologies, Inc.) were cultured in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and penicillin (100 U/ml)-streptomycin (100 mg/ml) at 37°C, with 5% CO 2 .

Techniques: Expressing, Western Blot

Journal: Oncology Letters

Article Title: Polyphyllin I and VII potentiate the chemosensitivity of A549/DDP cells to cisplatin by enhancing apoptosis, reversing EMT and suppressing the CIP2A/AKT/mTOR signaling axis

doi: 10.3892/ol.2019.10895

Figure Lengend Snippet: PPI and PPVII reverse EMT and suppress the CIP2A/AKT/mTOR pathway. A549/DDP cells were incubated with DDP (6 µg/ml), PPI (0.75 µg/ml), PPVII (0.75 µg/ml), or DDP (6 µg/ml) in combination with either PPI (0.75 µg/ml) or PPVII (0.75 µg/ml) for 24 h, then cells were harvested for further western blotting analysis. (A) The protein expression levels of E-cadherin, vimentin, α-SMA in A549/DDP cells were measured. (B) The protein expression levels of CIP2A, p-AKT, AKT, p-mTOR, m-TOR were measured. PPI, Polyphyllin I; PPVII, polyphyllin VII; DDP, cisplatin; p-AKT, phosphorylated-protein kinase B; mTOR, mammalian target of rapamycin; CIP2A, cancerous inhibitor of protein phosphatase 2A; SMA, smooth muscle actin; E, epithelial.

Article Snippet: Human NSCLC cell lines A549 and DDP-resistant A549/DDP cells (OriGene Technologies, Inc.) were cultured in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and penicillin (100 U/ml)-streptomycin (100 mg/ml) at 37°C, with 5% CO 2 .

Techniques: Incubation, Western Blot, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: Establishment of zebrafish NSCLC BM xenograft models. ( A ) Schematic diagram of imaging and collecting zebrafish brain. The zebrafish brain was imaged from vertical view (blue arrow) and excised (red dotted box) to extract RNA. ( B ) The schematic diagram showed zebrafish embryos at 2 dpf. The red dots indicated cancer cells injected into the perivitelline space (PVs). ( C ) Images of brain at different developmental stages of Tg ( fli-1 : EGFP) zebrafish (2-6 dpf). The white short dashed line indicated the midbrain of the zebrafish, and the white long dashed line indicated the zebrafish’s eyes and hindbrain. ( D ) The expression of claudin-5 in the zebrafish brain at different developmental stages (2-7 dpf). ( E ) The expression of mfsd2aa and mfsd2ab in the zebrafish brain at different developmental stages (2-6 dpf). ( F ) Survival curves of 2 dpf zebrafish with different tumor-bearing amount in PVs. About 50-400 H1975 cells (red fluorescence) were injected into the PVs of 2 dpf zebrafish, and the number of deaths was counted till 8 dpi. ( G - I ) Three human NSCLC cell lines were involved: H1975, A549 and H1299. About 100 cells were injected into the PVs of zebrafish at different developmental stages, and the brain of zebrafish was imaged at 1 dpi. The white arrows indicated cancer cells in blood vessels of zebrafish brain. ( J - K ) Quantification of the BM cells number in zebrafish at 1 dpi. Significance was considered when P values were lower than 0.05. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001. dpf: days post fertilization, dpi: days post injection

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: Imaging, Injection, Expressing, Fluorescence

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: NSCLC cells could not proliferate in the midbrain of zebrafish at 4 dpi. ( A ) The schematic diagram showed zebrafish embryos at 2 dpf. The red dots indicated cancer cells injected into the midbrain of zebrafish. ( B ) Survival curves of zebrafish with different tumor-bearing amount at midbrain. About 50-400 H1975 cells (red fluorescence) were injected into the midbrain of 2 dpf Tg ( fli-1 : EGFP) zebrafish, and the number of deaths was counted till 8 dpi. ( C - G ) Five human cancer cell lines were involved: human breast cancer cell lines MDA-MB-231 (positive control) and MCF-7 (negative control), human NSCLC cancer lines H1975, A549 and H1299. About 100 cells were injected into the midbrain of zebrafish at 2 dpf, and the midbrain of zebrafish was imaged at 1 dpi and 4 dpi. (H-M) Quantification of cell proliferation in the midbrain of zebrafish at 1 dpi and 4 dpi. The fold change of cells in the midbrain was determined by dividing the measured values at 1 dpi and 4 dpi by the average measured values at 1 dpi. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: Injection, Fluorescence, Positive Control, Negative Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: Zebrafish NSCLC BM xenograft models discriminated the BM potentials of different cell lines. ( A ) Transwell culture system was used to assess cell invasion ability in vitro , and five cancer cell lines were involved: MDA-MB-231, MCF-7, H1975, A549 and H1299. Invaded cells were stained with crystal violet (0.5 %) and imaged after 24 h incubation. ( B ) Quantification and comparison of invaded cells in vitro . Colonies were quantified using Image Pro Plus. ( C ) Five cancer cell lines were involved: MDA-MB-231, MCF-7, H1975, A549 and H1299. About 100 cells (red fluorescence) were injected into the PVs of Tg ( fli-1 : EGFP) zebrafish at 2 dpf, and the brain of zebrafish was imaged at 4 dpi. The white arrows indicated cancer cells in blood vessels of zebrafish brain. ( D ) Quantification of BM rate at 4 dpi. The number of BM cells in the same zebrafish at 4 dpi was divided by the number of BM cells at 1 dpi. The ratio of these two was named as BM potential. If the BM potential was greater than 1, it was considered that the zebrafish had brain metastasis. ( E - I ) Quantification of BM cells at 1 dpi and 4 dpi. The two dots connected by a straight line represented the number of BM cells of the same zebrafish at 1 dpi and 4 dpi. ( J ) Quantification and comparison of cell BM potential. The BM potential was determined by dividing the number of BM cells in the same zebrafish at 4 dpi by the number of BM cells at 1 dpi. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: In Vitro, Staining, Incubation, Comparison, Fluorescence, Injection

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: Zebrafish NSCLC BM xenograft models simultaneously discriminated the BM potentials of different cell lines. ( A ) The schematic diagram showed zebrafish embryos at 2 dpf. The red dots and green dots indicated cancer cells labeled with two different dyes were co-injected into the PVs of zebrafish. ( B ) About 50 H1975 cells (red fluorescence) and 50 H1975 cells (green fluorescence) were co-injected into the PVs of wild zebrafish at 2 dpf, and the brain of zebrafish was imaged at 4 dpi. The white long dashed line indicated the brain of zebrafish. ( C ) About 50 H1975 cells (red fluorescence) and 50 A549 cells (green fluorescence) were co-injected into the PVs of wild zebrafish at 2 dpf, and the brain of zebrafish was imaged at 4 dpi. ( D - G ) Quantification of BM cells in co-injection at 1 dpi and 4 dpi. The two dots connected by a straight line represented the number of BM cells of the same zebrafish at 1 dpi and 4 dpi. ( H and I ) The percentage of cells with two different dyes in the brain of zebrafish. The percentage of the cell line (red/green fluorescence) at 1 dpi was determined by dividing the number of BM cells (red/green fluorescence) by the total number of BM cells at 1 dpi. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: Labeling, Injection, Fluorescence

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: Expression of microRNA-330-3p (miR-330-3p) affected the NSCLC BM potential in zebrafish xenograft models. ( A and B ) Quantification of the miR-330-3p expression in vitro . The appropriate transfection concentration was determined between 1 nM and 50 nM, according to the instructions. ( C and D ) The expression of miR-330-3p in H1975 cell line was knocked down by anti-miR-330-3p inhibitor and the expression of miR-330-3p in A549 cell line was overexpressed by over-miR-330-3p mimics. Transwell culture system was used to evaluate the effectiveness of transfection in vitro . Invaded cells were stained with crystal violet (0.5 %) and imaged after 24 h incubation. ( E and F ) Quantification of invaded cells in vitro . Colonies were quantified using Image Pro Plus. ( G and H ) H1975, H1975 with under-expressed miR-330-3p, A549 and A549 with over-expressed miR-330-3p were involved. About 100 cells (red fluorescence) were injected into the PVs of Tg ( fli-1 : EGFP) zebrafish at 2 dpf to evaluate the effectiveness of transfection in vivo , and the brain of zebrafish was imaged at 4 dpi. The white arrows indicated cancer cells in blood vessels of zebrafish brain. ( I - L ) Quantification of BM cells at 1 dpi and 4 dpi. The two dots connected by a straight line represented the number of BM cells of the same zebrafish at 1 dpi and 4 dpi. ( M and N ) Quantification and comparison of the BM potentials of the transfected cell lines and the original cell lines. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: Expressing, In Vitro, Transfection, Concentration Assay, Staining, Incubation, Fluorescence, Injection, In Vivo, Comparison

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Zebrafish xenograft model for studying mechanism and treatment of non-small cell lung cancer brain metastasis

doi: 10.1186/s13046-021-02173-5

Figure Lengend Snippet: Zebrafish NSCLC BM xenograft models discriminated different chemosensitivities within 4 days. ( A and B ) NSCLC cell lines H1975 and A549 were involved. Transwell culture system was used to evaluate the effectiveness of osimertinib (1 µM) and gefitinib (2 µM) against cell invasion in vitro . Invaded cells were stained with crystal violet (0.5 %) and imaged after 24 h incubation. ( C and D ) Quantification of invaded cells in vitro. Colonies were quantified using Image Pro Plus. ( E and F ) NSCLC cell lines H1975 and A549 were involved. Tg ( fli-1 : EGFP) zebrafish embryos were injected about 100 cells (red fluorescence) into the PVs at 2 dpf and administered with osimertinib (1 µM) and gefitinib (13 µM) by intracardiac injection. ( G and H ) Quantification of cell brain metastasis at 4 dpi. Fold change of BM cell number was determined by dividing the number of BM cells in the same zebrafish at 4 dpi by the number of BM cells at 1 dpi. (ns) indicated statistical insignificance, (*) indicated statistical significance P < 0.05, (**) P < 0.01 and (***) P < 0.001

Article Snippet: Human NSCLC lung cancer cell lines, A549, H1975, H1299, originally from American Type Culture Collection, were cultured in accordance with standard requirements.

Techniques: In Vitro, Staining, Incubation, Injection, Fluorescence

Journal: Cancer Research

Article Title: Regression of Melanoma in a Murine Model by RLIP76 Depletion

doi: 10.1158/0008-5472.can-05-3534

Figure Lengend Snippet: Figure 1. Comparison of RLIP76 levels in cultured malignant cells versus nonmalignant cells. Aliquots of crude detergent extracts of the membrane fractions of malignant cells (H1618, H358, OVCAR-3, PC-3, MCF-7, B16, HepG2, and DG-1) and nonmalignant cells (HAVSM, HUVEC, HLMVEC, and HLBEC), containing 200 Ag protein were used for SDS-PAGE and Western blotting against anti-RLIP76 IgG as primary antibody and horseradish peroxidase–conjugated goat anti-rabbit IgG as secondary antibody and developed with 4-chloro-1-napthol as chromogenic substrate. Results were quantified by scanning densitometry of the full-length RLIP76 protein band near 109 kDa. h-Actin was used as an internal control.

Article Snippet: Human small cell lung cancer (SCLC) lines H1618 and non–small cell lung cancer (NSCLC) lines H358 (bronchioalveolar), PC-3 (human prostate), and B16 mouse skin melanoma were purchased from American Type Culture Collection (Manassas, VA).

Techniques: Comparison, Cell Culture, Membrane, SDS Page, Western Blot, Control

Journal: Journal of medicinal chemistry

Article Title: A Potent and Highly Efficacious Bcl-2/Bcl-xL Inhibitor

doi: 10.1021/jm4001105

Figure Lengend Snippet: Antitumor activity of compounds 31 and 32 in the H146 small-cell lung cancer xenograft model in SCID mice. Tumors were grown to an average size of 126 mm3 and compound 31 or 32 was administered at 15 mg/kg intravenously, daily, 5 days a week for 2 weeks. (a). Tumor growth. (b). Animal body weight.

Article Snippet: 7 Human small cell lung cancer cell lines H146, H1963, H187, and H1417 were purchased from American Type Culture Collection (ATCC) and were maintained in RPMI-1640 medium containing 10% FBS.

Techniques: Activity Assay

Journal: Molecular Medicine

Article Title: Deficiency of mindin reduces renal injury after ischemia reperfusion

doi: 10.1186/s10020-022-00578-2

Figure Lengend Snippet: Mindin expression was elevated in renal IR injury in vivo and in vitro. A Mindin measured by western blot analysis in sham mice and IR mice. Quantitative analysis of mindn protein expression. Expression levels were normalized to GAPDH and are presented relative to the sham-operated kidney. *P < 0.05 versus Sham. B Representative photomicrographs showing IHC analysis for mindin after 30 min renal ischemia and 24 h reperfusion (magnification 400 ×). C Mindin measured by western blot analysis in HK-2 cells after 24 h hypoxia and 2, 4, and 8 h reoxygenation. Quantitative analysis of mindin protein expression. Expression levels were normalized to GAPDH and are presented relative to the control cells. *P < 0.05 versus Control. D Representative photomicrographs showing immunofluorescence staining for mindin after 24 h hypoxia and 8 h reoxygenation (magnification 400 ×)

Article Snippet: Mindin protein (HY-P70142, MedChemExpress) and TLR4 protein (HY-P73586, MedChemExpress) were used to study the protein binding in CM5 electronic chips by biomolecular interaction instrument (Biacore T2000).

Techniques: Expressing, In Vivo, In Vitro, Western Blot, Control, Immunofluorescence, Staining

Journal: Molecular Medicine

Article Title: Deficiency of mindin reduces renal injury after ischemia reperfusion

doi: 10.1186/s10020-022-00578-2

Figure Lengend Snippet: Mindin deficiency attenuated renal IR injury. A Mindin protein expression in kidneys from WT and Mindin KO mice with and without IR. B Levels of BUN and Cre after renal IR injury in WT and mindin KO mice. *P < 0.05 versus WT Sham or KO Sham. # P < 0.05 versus WT IR. C Representative photomicrographs showing HE staining in WT and mindin KO mice with or without renal IR injury (magnification 400 ×). D Degree of renal damage graded using the acute tubular necrosis (ATN) score. *P < 0.05 versus WT Sham or KO Sham. # P < 0.05 versus WT IR

Article Snippet: Mindin protein (HY-P70142, MedChemExpress) and TLR4 protein (HY-P73586, MedChemExpress) were used to study the protein binding in CM5 electronic chips by biomolecular interaction instrument (Biacore T2000).

Techniques: Expressing, Staining

Journal: Molecular Medicine

Article Title: Deficiency of mindin reduces renal injury after ischemia reperfusion

doi: 10.1186/s10020-022-00578-2

Figure Lengend Snippet: Mindin deficiency inhibited TLR4/JNK/NF-κB signaling activation in renal IR injury A TLR4, phosphorylated JNK, phosphorylated P65, and phosphorylated IκBα measured by western blot analysis in kidneys of WT and mindin KO mice induced by IR or not. B – E Quantitative analysis of TLR4, phosphorylated JNK, phosphorylated P65, and phosphorylated IκBα protein expression. *P < 0.05 versus WT Sham or KO Sham. # P < 0.05 versus WT IR

Article Snippet: Mindin protein (HY-P70142, MedChemExpress) and TLR4 protein (HY-P73586, MedChemExpress) were used to study the protein binding in CM5 electronic chips by biomolecular interaction instrument (Biacore T2000).

Techniques: Activation Assay, Western Blot, Expressing

Journal: Molecular Medicine

Article Title: Deficiency of mindin reduces renal injury after ischemia reperfusion

doi: 10.1186/s10020-022-00578-2

Figure Lengend Snippet: Mindin overexpression promoted production of inflammatory mediators and activation of TLR4/JNK/ NF-κB signaling after HR in vitro. A Representative photomicrographs showing the immunofluorescence of GFP in adGFP and adMindin cells. B Mindin protein expression in adGFP and adMindin cells. C – E Protein levels of TNF-α and MCP-1 in the media of cultured TECs by Western blot. *P < 0.05 versus adGFP Con or adMindn Con. # P < 0.05 versus adGFP HR. F TLR4, phosphorylated JNK, phosphorylated P65, and phosphorylated IκBα measured by western blot analysis in adGFP and adMindin cells induced by HR or not. G – J Quantitative analysis of TLR4, phosphorylated JNK, phosphorylated P65, and phosphorylated IκBα protein expression. *P < 0.05 versus adGFP Con or adMindn Con. # P < 0.05 versus adGFP HR. K Quantitative analysis of TLR4, JNK, P65, TNF-α and MCP-1 mRNA expression in mindin-overexpressed HK-2 cells. ***P < 0.001 versus adGFP HR group

Article Snippet: Mindin protein (HY-P70142, MedChemExpress) and TLR4 protein (HY-P73586, MedChemExpress) were used to study the protein binding in CM5 electronic chips by biomolecular interaction instrument (Biacore T2000).

Techniques: Over Expression, Activation Assay, In Vitro, Immunofluorescence, Expressing, Cell Culture, Western Blot

Journal: Molecular Medicine

Article Title: Deficiency of mindin reduces renal injury after ischemia reperfusion

doi: 10.1186/s10020-022-00578-2

Figure Lengend Snippet: Mindin knockdown inhibited the activation of TLR4/JNK/ NF-κB signaling after HR in vitro. A TLR4, Mindin, phosphorylated JNK, phosphorylated P65, and phosphorylated IκBα measured by western blot analysis in si-Mindin, si-TLR4 and si-JNK cells induced by HR respectively. B – F Quantitative analysis of TLR4, Mindin, phosphorylated JNK, phosphorylated P65, and phosphorylated IκBα protein expression. * P < 0.05, ** P < 0.01 versus HR. G Quantitative analysis of TLR4, JNK, P65, TNF-α and MCP-1 mRNA expression in si-Mindin, si-TLR4 and si-JNK cells induced by HR respectively. ***P < 0.001 versus control HR group

Article Snippet: Mindin protein (HY-P70142, MedChemExpress) and TLR4 protein (HY-P73586, MedChemExpress) were used to study the protein binding in CM5 electronic chips by biomolecular interaction instrument (Biacore T2000).

Techniques: Knockdown, Activation Assay, In Vitro, Western Blot, Expressing, Control

Journal: Molecular Medicine

Article Title: Deficiency of mindin reduces renal injury after ischemia reperfusion

doi: 10.1186/s10020-022-00578-2

Figure Lengend Snippet: Co-immunoprecipitation of TLR4 and mindin in HK-2 cells. Lysates of HK-2 cells were incubated with anti-TLR4 antibody and immune complexes were precipitated by protein A/G beads. The TLR4 complex was determined TLR4 and mindin proteins in the complex. The whole cell lysate was used to detect TLR4 and mindin by Western blot

Article Snippet: Mindin protein (HY-P70142, MedChemExpress) and TLR4 protein (HY-P73586, MedChemExpress) were used to study the protein binding in CM5 electronic chips by biomolecular interaction instrument (Biacore T2000).

Techniques: Immunoprecipitation, Incubation, Western Blot

Journal: Molecular Medicine

Article Title: Deficiency of mindin reduces renal injury after ischemia reperfusion

doi: 10.1186/s10020-022-00578-2

Figure Lengend Snippet: Binding assy of mindin to TLR4 proteins by SPR. Different concen-trations of TLR4 protein (3.125, 6.25, 12.5, 25.0, 50.0, 100.0 nM) were performed to analyse the binding and dissociation rate constants between mindin and TLR4 proteins. The binding constant of them was calculated to evaluate protein binding

Article Snippet: Mindin protein (HY-P70142, MedChemExpress) and TLR4 protein (HY-P73586, MedChemExpress) were used to study the protein binding in CM5 electronic chips by biomolecular interaction instrument (Biacore T2000).

Techniques: Binding Assay, Protein Binding

Journal: Molecular Medicine

Article Title: Deficiency of mindin reduces renal injury after ischemia reperfusion

doi: 10.1186/s10020-022-00578-2

Figure Lengend Snippet: Mindin protein was expressed massively in renal tubule tissues of human hydronephrosis patients by immunohistochemistry. Red arrow indicate mindin expression

Article Snippet: Mindin protein (HY-P70142, MedChemExpress) and TLR4 protein (HY-P73586, MedChemExpress) were used to study the protein binding in CM5 electronic chips by biomolecular interaction instrument (Biacore T2000).

Techniques: Immunohistochemistry, Expressing