Journal: Cell Death Discovery

Article Title: Trifluoperazine causes mast cell apoptosis through a secretory granule-mediated pathway

doi: 10.1038/s41420-026-03122-x

Figure Lengend Snippet: A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway epithelial cells (HSAECs) were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.

Article Snippet: Primary human lung fibroblasts (HLFs) (PCS-201-013) and primary human lung smooth muscle cells (HLSMCs)(PCS-130-010) were obtained from American Type Culture Collection (ATCC, Manassas, VA) and cultured following the manufacturer’s instructions.

Techniques: Staining, Derivative Assay, Control, Agarose Gel Electrophoresis, Marker

Journal: Frontiers in immunology

Article Title: Heterozygous premature termination in zinc-finger domain of Krüppel-like factor 2 gene associates with dysregulated immunity.

doi: 10.3389/fimmu.2022.819929

Figure Lengend Snippet: FIGURE 1 (A) Family tree illustrates the individuals with KLF2 c.951dup p.(Glu318Argfs*87) mutation. Black arrows point at the index patient IV:2 diagnosed with idiopathic juvenile arthritis and the index patient III:2 with lymphopenia of unknown origin. The variant positive family members suffer from lymphopenia, arthritis, lymphoma, and malignancies marked with small black boxes in four alternative corners, each indicating a different condition. Summary and timeframe (years) of clinical events in patients II:1 (B), II:2 (C), III:2 (D) and IV:2 (E) is shown. The major health events and medications are indicated by arrows (a to x). The corresponding health events are explained in detail in chapter 2.1. CLL, Chronic lymphocytic leukemia; MTX, methotrexate; SSZ, sulfasalazine; CSA, cyclosporine; PL, prednisolone; ATM, sodium aurothiomalate; CP, cyclophosphamide; CMB, chlorambucil; VS, vincristine sulfate; H, hydroxydaunorubicin; CVID, common variable immunodeficiency; HCQS, hydroxychloroquine; ETN, etanercept.

Article Snippet: After permeabilization the nuclei were washed and stained for APC conjugated antibody against human KLF2 (Miltenyi Biotec; 130-111-040) along with isotype control (Miltenyi Biotec; 130- 120-709) for 30 minutes at 4°C.

Techniques: Mutagenesis, Variant Assay, Medications

Journal: Frontiers in immunology

Article Title: Heterozygous premature termination in zinc-finger domain of Krüppel-like factor 2 gene associates with dysregulated immunity.

doi: 10.3389/fimmu.2022.819929

Figure Lengend Snippet: FIGURE 2 (A) KLF2 protein structure. The conserved C2H2-type zinc finger structure at the C-terminus consists of three loops that are crucial for DNA- binding. The KLF2 c.951dup p.(Glu318Argfs*87) mutation is located in the middle of the second zinc finger loop. The duplication of Glu318 causes a shift in the reading frame which leads to a stop codon 87 amino acids later. (B) Comparison of the sequence coding the human mutant to human, mouse, and zebrafish with KLF2 zinc fingers. The mutated part of the KLF2 sequence [c.951dup p.(Glu318Argfs*87)] is typed with red color. The sequence shown begins with the nuclear localization signal (NLS: amino acids (aa) 254 - 274), followed by the first zinc- finger (Zf1: aa277 - 297), the second zinc-finger (Zf2: aa304 - 327) and the third zinc finger (Zf3: aa334 - 254). Within the zinc finger sequences, there are three conserved DNA-binding domains (marked with blue boxes). The black arrows point at important amino acids that bind directly to DNA. The KLF2-PE/APC antibody binding sequences (A) are intact both in the wildtype and the variant alleles. KLF2 was detected by western blot in patient (III:2) skin fibroblasts (C) and neutrophils (D) as 37 kDa and 60 kDa molecular weight forms correspondingly. 30µg of protein was separated on a 12% gel and transferred into PVDF membrane. The KLF2 and GAPDH antibody binding was detected by anti-mouse or anti- rabbit secondary antibodies (IRDye 680RD or 800CW). The membranes were imaged by Odyssey infrared scanner and band intensities quantified from three different sample sets. (E) KLF2 expression in CD4+ and CD8+ lymphocytes. The KLF2 expression of isolated PBMCs from mutation carriers (III:1 and III:2) and sex-matched controls was determined by flow cytometry. (F) Localization of KLF2 protein was studied in control PBMCs with flow cytometry using three different permeabilization strategies: first without any permeabilization/fixation (used when staining cell membrane targets), second with Cytoperm buffer (used when staining cytoplasmic targets) and finally, a Fixation/permeabilization buffer (used when staining nuclear transcription factors). Quantitation of MFI values is shown in bar chart below. (G) Nuclei from patient and control PBMCs were isolated, fixed, permeabilized and stained with KLF-APC and isotype control antibodies. Quantitation of MFI values is shown in bar chart below.

Article Snippet: After permeabilization the nuclei were washed and stained for APC conjugated antibody against human KLF2 (Miltenyi Biotec; 130-111-040) along with isotype control (Miltenyi Biotec; 130- 120-709) for 30 minutes at 4°C.

Techniques: Binding Assay, Mutagenesis, Comparison, Sequencing, Variant Assay, Western Blot, Molecular Weight, Membrane, Expressing, Isolation, Cytometry, Control, Staining, Quantitation Assay

Journal: Frontiers in immunology

Article Title: Heterozygous premature termination in zinc-finger domain of Krüppel-like factor 2 gene associates with dysregulated immunity.

doi: 10.3389/fimmu.2022.819929

Figure Lengend Snippet: FIGURE 3 KLF2 interactome analysis. (A) Schematic workflow overview of the MAC-tagged based BioID purification coupled with mass spectrometry. (B) Dot-plot visualization (BFDR ≤0.05) of the KLF2-WT and KLF2-MUT interactors (prohits-viz.org). Each node corresponds to the abundance of the average spectral count for each prey. (C, D) The polar plot shows the molecular level localization of KLF2-WT and KLF2-MUT obtained by MS-microscopy respectively (proteomics.fi). ACTF, Actin filament; JUNC, Cell junction; CEN, Centrosome; CHR, Chromatin; ER, Endoplasmic reticulum; END, Endosome; EXO, Exosome; FOCA, Focal adhesion; GOL, Golgi; INTF, Intermediate filament; LYS, Lysosome; MIC, Microtubule; MT, Mitochondrion; NE, Nuclear envelope; NUC, Nucleolus; NUCPL, Nucleoplasm; PER, Peroxisome; PM, Plasmamembrane; PRO, Proteasome. (E, F) Significantly enriched (q < 0.05, calculated with Fisher exact test and Benjamini–Hochberg multiple-testing correction) Reactome pathway annotations from the KLF2-WT and KLF2-MUT interactors, respectively. Y-axis corresponds to the percentage of the interactors found in the given pathway.

Article Snippet: After permeabilization the nuclei were washed and stained for APC conjugated antibody against human KLF2 (Miltenyi Biotec; 130-111-040) along with isotype control (Miltenyi Biotec; 130- 120-709) for 30 minutes at 4°C.

Techniques: Mass Spectrometry, Microscopy

Journal: Frontiers in immunology

Article Title: Heterozygous premature termination in zinc-finger domain of Krüppel-like factor 2 gene associates with dysregulated immunity.

doi: 10.3389/fimmu.2022.819929

Figure Lengend Snippet: FIGURE 5 (A) CD3+CD4+ T cell subgroups in KLF2 c.951dup p.(Glu318Argfs*87) variant positive family members compared to healthy controls. Mutation carriers´ and controls´ unstimulated PBMCs were stained for regulatory T helper cell markers CD4, CD25, CD127 and FOXP3 and analyzed by flow cytometry. Th1 and Th 17 subpopulations were stained ex vivo (whole blood) with CD4, CXCR3, CCR6, CD45RA and CD10 antibodies and analyzed by flow cytometry. (B) Representative results of flow cytometry analysis of CD25+CD127lowFOXP3+ regulatory T cells (Treg). The gating strategy is presented along with the data. Regulatory T cells of patient (III:2) and sex-matched controls´ were studied for the expression of CD45RA, CCR6, CCR7 and CXCR3 with flow cytometry. (C) The Treg populations I-V were studied for the expression of CCR6, CCR7 and CXCR3. Representative results of patient and control Treg populations are shown. The regulatory T cell populations I, II, III, IV and V are presented as the number of cells and percentages. The percentage of CXCR3, CCR6 and CCR7 positive cells.

Article Snippet: After permeabilization the nuclei were washed and stained for APC conjugated antibody against human KLF2 (Miltenyi Biotec; 130-111-040) along with isotype control (Miltenyi Biotec; 130- 120-709) for 30 minutes at 4°C.

Techniques: Variant Assay, Mutagenesis, Staining, Cytometry, Ex Vivo, Expressing, Control

Journal: The Journal of Biological Chemistry

Article Title: TGF-β/SMAD3 Pathway Stimulates Sphingosine-1 Phosphate Receptor 3 Expression

doi: 10.1074/jbc.M116.740084

Figure Lengend Snippet: Up-regulation of S1PR3 in human lung adenocarcinomas. A, qPCR quantitation of S1PR3 mRNA in cDNA arrays of human lung adenocarcinoma specimens (OriGene, HLRT101 and HLRT105). **, p < 0.01, Student's t test. B, qPCR quantitation of S1PR2 mRNA in a cDNA array of human lung cancers (OriGene, HLRT105). **, p < 0.01, Student's t test. C, HEK293 cells were transfected with S1PR3 or pcDNA vector. Transfected cells were immunostained with anti-S1PR3 (Cayman Chemical) (IMF, left panels). Arrows, nonspecific fluorescent precipitates used for image orientation. Scale bar = 33 μm. D, anti-S1PR3 staining of human lung adenocarcinoma tumor microarray (Accumax 306). AdC, adenocarcinoma; N, adjacent normal lung tissue. E, immunostaining intensity was quantitated with the National Institutes of Health ImageJ software. Data, analyzed with GraphPad Prism 5 software, are shown as mean ± S.E. Statistical significance was analyzed by Student's t test. F, representative images of anti-S1PR3 staining of human lung adenocarcinoma and the respective adjacent normal lung epithelial tissue. G, quantitation of anti-S1PR3 staining of human lung squamous carcinoma microarray (Accumax 306). Data are mean ± S.E. Statistical significance was analyzed by Student's t test. H, representative images of anti-S1PR3 staining of human lung squamous carcinoma and the respective adjacent normal lung epithelial tissue.

Article Snippet: The pathological relevance of this in vitro observation was investigated by measuring mRNA levels of S1PR3 in cDNA microarrays of human lung adenocarcinoma specimens (OriGene, HLRT).

Techniques: Quantitation Assay, Transfection, Plasmid Preparation, Staining, Microarray, Immunostaining, Software

Journal: The Journal of Biological Chemistry

Article Title: TGF-β/SMAD3 Pathway Stimulates Sphingosine-1 Phosphate Receptor 3 Expression

doi: 10.1074/jbc.M116.740084

Figure Lengend Snippet: Oncogenic K-Ras mutant stimulates S1PR3 expression. A, LSL-K-RasG12D mice were intratracheally injected with empty adenoviral (Ad-Ctrl) or Ad-Cre particles (1 × 108 pfu). The development of lung adenocarcinomas (arrows) was analyzed 2 months later. Scale bar = 0.5 cm. B, K-RasG12D mice were injected with Ad-Ctrl or Ad-Cre particles. 2 months later, levels of S1PRs in lungs were measured by qPCR analysis. ** and *, p < 0.01 and 0.05, respectively. NS, non-statistically significant. n = 5, Student's t test. C, immunohistochemical staining of S1PR3 in lung specimens from wild-type or K-Ras transgenic mice. Note that levels of S1PR3 are profoundly increased in lung adenocarcinoma of K-Ras transgenic mice (arrows). Scale bar = 200 μm.

Article Snippet: The pathological relevance of this in vitro observation was investigated by measuring mRNA levels of S1PR3 in cDNA microarrays of human lung adenocarcinoma specimens (OriGene, HLRT).

Techniques: Mutagenesis, Expressing, Injection, Immunohistochemical staining, Staining, Transgenic 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: 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: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 5. RAGE immunoblotting analysis of human intestine. SDS-PAGE of lung lysate (1.5 μg/lane) as well as lysates from small intestine (SI) and colon (50 μg/lane) was performed under reducing conditions. Membranes were stained with three different anti-RAGE antibodies; SC A11 (A), AF1145 (B), and AB D01P (C). Membranes were also stained with the respective secondary antibodies only (negative control). GAPDH was used as a loading control. The uncropped images are shown in the supplementary material of the manuscript.

Article Snippet: Human Lung Whole Tissue Lysate (Adult Whole Normal) was obtained from Novus Biologicals and protein extracts from healthy human small intestine and colon were obtained from Santa Cruz Biotechnology.

Techniques: Western Blot, SDS Page, Staining, Negative Control, Control

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 4. (A) RAGE immunoblotting analysis of human lung lysate. SDS-PAGE of lung lysate (1 μg/lane; PNGase F -) and deglycosylated lung lysate (1 μg/lane; PNGase F +) was performed under reducing conditions. Membranes were stained with four different anti-RAGE antibodies. Membranes were also stained with the respective secondary antibodies only (negative control). GAPDH was used as a loading control. (B) RAGE immunoblotting analysis of RAGE overexpressing (+) and control (−) HEK293T cells. SDS-PAGE was performed under reducing conditions. For WB analysis membranes were stained with anti-FLAG and anti-RAGE (AB D01P) antibodies. The uncropped images are shown in the supplementary material of the manuscript.

Article Snippet: Human Lung Whole Tissue Lysate (Adult Whole Normal) was obtained from Novus Biologicals and protein extracts from healthy human small intestine and colon were obtained from Santa Cruz Biotechnology.

Techniques: Western Blot, SDS Page, Staining, Negative Control, Control

Journal: Heliyon

Article Title: Human intestine and placenta exhibit tissue-specific expression of RAGE isoforms.

doi: 10.1016/j.heliyon.2023.e18247

Figure Lengend Snippet: Fig. 6. RAGE immunoblotting analysis of human placenta derived from different healthy (H) as well as GDM-, FGR- and PE-affected pregnancies. SDS-PAGE of lung lysate (1 μg/lane) as well as lysates from the placentas (25 μg/lane) was performed under reducing conditions. For WB analysis membranes were stained with three different anti-RAGE antibodies; SC A11 (A), AF1145 (B), and AB D01P (C). Membranes were also stained with the respective secondary antibodies only (negative control). Ponceau S staining was used as a loading control. The uncropped images are shown in the supplementary material of the manuscript.

Article Snippet: Human Lung Whole Tissue Lysate (Adult Whole Normal) was obtained from Novus Biologicals and protein extracts from healthy human small intestine and colon were obtained from Santa Cruz Biotechnology.

Techniques: Western Blot, Derivative Assay, SDS Page, Staining, Negative Control, Control

Journal: Oncotarget

Article Title: MicroRNA-224 is implicated in lung cancer pathogenesis through targeting caspase-3 and caspase-7.

doi: 10.18632/oncotarget.5224

Figure Lengend Snippet: Figure 2: The expressions of CASP3 and CASP7 mRNA are down regulated in NSCLC. a. CASP7 mRNA expressions in thirty paired NSCLC tissues and NATs. b. CASP3 mRNA expressions in thirty paired NSCLC tissues and NATs. GAPAH was used for normalization. c. and d. Kaplan-Meier plots of overall survival of lung cancer patients, stratified by expression of CASP7 (1926 patients) c. or CASP3 (1926 patients) d.. Data obtained from the Kaplan-Meier plotter database (kmplot.com/analysis).

Article Snippet: nsclc tissue samples Human Lung Tissue Microarrays (IMH-358) were purchased from Novus Biologicals, San Diego, CA.

Techniques: Expressing

Journal: Oncotarget

Article Title: MicroRNA-224 is implicated in lung cancer pathogenesis through targeting caspase-3 and caspase-7.

doi: 10.18632/oncotarget.5224

Figure Lengend Snippet: Figure 4: Expressions of CASP3 and CASP7 are inversely correlated with miR-224. a. Representative pictures and summary of co-expression analyses for miR-224 and CASP7 in NSCLC tissues. MiR-224 was detected by using 5’-DIG labeled LNA probe (purple) and CASP7 was detected by immunohistochemistry (red). Left: High CASP7 and low/neg miR-224, Middle: similar expression of CASP7 and miR-224, right: Low/neg CASP7 and high miR-224. b. Representative pictures and summary of co-expression analyses for miR- 224 and CASP3 in NSCLC tissues. MiR-224 was detected by using 5’-DIG labeled LNA probe (purple) and CASP3 was detected by immunohistochemistry (red). Left: High CASP3 and low/neg miR-224, Middle: similar expression of CASP3 and miR-224, right: Low/ neg CASP3 and high miR-224. c. In the cases of NSCLC where both miR-224 and CASP7 expression were noted, no detectable CASP7 was found in cancer cells overexpressing miR-224 (purple arrow).

Article Snippet: nsclc tissue samples Human Lung Tissue Microarrays (IMH-358) were purchased from Novus Biologicals, San Diego, CA.

Techniques: Expressing, Labeling, Immunohistochemistry