Different subcellular localizations and functions of Arabidopsis myosin VIII-6

Ocal microscopy. A. Root of 5-day-old seedling. Scale bar: 50 μm. B. Lateral root of 20-day-old seedling. Scale bar: 20 μm. C(1) and C(2). Two images of the same 20-day-old seedling root. C(1) shows the root cap, scale bar: 20 μm, and C(2) shows the upper part. Scale bar: 50 μm. A similar pattern of GFP-ATM1 localization is seen in all roots: diffuse at the root cap, then dots, then more polarized organization along the transverse sides. D. GFP-ATM1 in root hair, scale bar: 10 μm. Arrows show the direction of the root caps.<p><b>Copyright information:</b></p><p>Taken from "Different subcellular localizations and functions of Arabidopsis myosin VIII"</p><p>http://www.biomedcentral.com/1471-2229/8/3</p><p>BMC Plant Biology 2008;8():3-3.</p><p>Published online 8 Jan 2008</p><p>PMCID:PMC2275265.</p><p></p

Different subcellular localizations and functions of Arabidopsis myosin VIII-0

P starting from the ATG and a reverse primer corresponding to the 3' end of ATM1 including its stop codon. The size of the expected fragment was 1734 bp. The template DNA was as follows: Lane 1. DNA from transgenic plants expressing GFP-ATM1(IQ-tail). Lane 2. DNA from wt plants. Lane 3. DNA from the plasmid used to generate the transgenic plants. Lane 4. Molecular weight markers. B. Western blot analysis showing sizes and levels of the expressed transgenes: Lane 1. GFP alone. Lane 2. GFP-ATM1(IQ-tail). Detection was performed with anti-GFP antibody.<p><b>Copyright information:</b></p><p>Taken from "Different subcellular localizations and functions of Arabidopsis myosin VIII"</p><p>http://www.biomedcentral.com/1471-2229/8/3</p><p>BMC Plant Biology 2008;8():3-3.</p><p>Published online 8 Jan 2008</p><p>PMCID:PMC2275265.</p><p></p

Different subcellular localizations and functions of Arabidopsis myosin VIII-2

D with a confocal microscope. A-B. Cells with GFP-ATM1(IQ-tail) organized in dots (arrows). A. GFP-ATM1. B. BFA bodies formed in these cells, shown by FM4-64. Note that the dotted pattern is not disrupted by the treatment (arrows). C. Overlay of A and B. Scale bar 10 μm. D-F Showing cells near the root cap where ATM1 is found in BFA bodies. D. GFP-ATM1, E. BFA bodies stained by FM4-64, F. overlay of D and E. Scale bar 10 μm. All images in this figure are composed of one optic section.<p><b>Copyright information:</b></p><p>Taken from "Different subcellular localizations and functions of Arabidopsis myosin VIII"</p><p>http://www.biomedcentral.com/1471-2229/8/3</p><p>BMC Plant Biology 2008;8():3-3.</p><p>Published online 8 Jan 2008</p><p>PMCID:PMC2275265.</p><p></p

Different subcellular localizations and functions of Arabidopsis myosin VIII-4

ATM1(IQ-tail) (dots of 980 ± 145 nm in diameter). B. FM4-64. C. Overlay of A and B (1 optic section). D. GFP-ATM1(IQ-tail) (dots of 630 ± 60 nm). E. FYVE-DsRED. F. Overlay of D and E (1 optic section). G. RFP-ATM1(IQ-tail) (dots of 300 ± 100, colored green for ease of demonstration). H. GFP-ARA7 (colored magenta for ease of demonstration). I. Overlay of G and H (1 optic section). J. RFP-ATM1(IQ-tail) (dots of 570 ± 75 nm, colored green for ease of demonstration). K. ARA6-GFP (colored magenta for ease of demonstration). L. Overlay of J. and K (1 optic section). Arrows show co-localization. Scale bars 5 μm. The microscope focus in A-I was in the cytoplasm while the focus in J-L was on the plasma membrane.<p><b>Copyright information:</b></p><p>Taken from "Different subcellular localizations and functions of Arabidopsis myosin VIII"</p><p>http://www.biomedcentral.com/1471-2229/8/3</p><p>BMC Plant Biology 2008;8():3-3.</p><p>Published online 8 Jan 2008</p><p>PMCID:PMC2275265.</p><p></p

Different subcellular localizations and functions of Arabidopsis myosin VIII-5

P starting from the ATG and a reverse primer corresponding to the 3' end of ATM1 including its stop codon. The size of the expected fragment was 1734 bp. The template DNA was as follows: Lane 1. DNA from transgenic plants expressing GFP-ATM1(IQ-tail). Lane 2. DNA from wt plants. Lane 3. DNA from the plasmid used to generate the transgenic plants. Lane 4. Molecular weight markers. B. Western blot analysis showing sizes and levels of the expressed transgenes: Lane 1. GFP alone. Lane 2. GFP-ATM1(IQ-tail). Detection was performed with anti-GFP antibody.<p><b>Copyright information:</b></p><p>Taken from "Different subcellular localizations and functions of Arabidopsis myosin VIII"</p><p>http://www.biomedcentral.com/1471-2229/8/3</p><p>BMC Plant Biology 2008;8():3-3.</p><p>Published online 8 Jan 2008</p><p>PMCID:PMC2275265.</p><p></p

DataSheet1_Integrated transcriptome and cell phenotype analysis suggest involvement of PARP1 cleavage, Hippo/Wnt, TGF-β and MAPK signaling pathways in ovarian cancer cells response to cannabis and PARP1 inhibitor treatment.docx

Introduction:Cannabis sativa is utilized mainly for palliative care worldwide. Ovarian cancer (OC) is a lethal gynecologic cancer. A particular cannabis extract fraction ('F7′) and the Poly(ADP-Ribose) Polymerase 1 (PARP1) inhibitor niraparib act synergistically to promote OC cell apoptosis. Here we identified genetic pathways that are altered by the synergistic treatment in OC cell lines Caov3 and OVCAR3.Materials and methods: Gene expression profiles were determined by RNA sequencing and quantitative PCR. Microscopy was used to determine actin arrangement, a scratch assay to determine cell migration and flow cytometry to determine apoptosis, cell cycle and aldehyde dehydrogenase (ALDH) activity. Western blotting was used to determine protein levels.Results: Gene expression results suggested variations in gene expression between the two cell lines examined. Multiple genetic pathways, including Hippo/Wnt, TGF-β/Activin and MAPK were enriched with genes differentially expressed by niraparib and/or F7 treatments in both cell lines. Niraparib + F7 treatment led to cell cycle arrest and endoplasmic reticulum (ER) stress, inhibited cell migration, reduced the % of ALDH positive cells in the population and enhanced PARP1 cleavage.Conclusion: The synergistic effect of the niraparib + F7 may result from the treatment affecting multiple genetic pathways involving cell death and reducing mesenchymal characteristics.</p

Table1_Integrated transcriptome and cell phenotype analysis suggest involvement of PARP1 cleavage, Hippo/Wnt, TGF-β and MAPK signaling pathways in ovarian cancer cells response to cannabis and PARP1 inhibitor treatment.XLSX

Introduction:Cannabis sativa is utilized mainly for palliative care worldwide. Ovarian cancer (OC) is a lethal gynecologic cancer. A particular cannabis extract fraction ('F7′) and the Poly(ADP-Ribose) Polymerase 1 (PARP1) inhibitor niraparib act synergistically to promote OC cell apoptosis. Here we identified genetic pathways that are altered by the synergistic treatment in OC cell lines Caov3 and OVCAR3.Materials and methods: Gene expression profiles were determined by RNA sequencing and quantitative PCR. Microscopy was used to determine actin arrangement, a scratch assay to determine cell migration and flow cytometry to determine apoptosis, cell cycle and aldehyde dehydrogenase (ALDH) activity. Western blotting was used to determine protein levels.Results: Gene expression results suggested variations in gene expression between the two cell lines examined. Multiple genetic pathways, including Hippo/Wnt, TGF-β/Activin and MAPK were enriched with genes differentially expressed by niraparib and/or F7 treatments in both cell lines. Niraparib + F7 treatment led to cell cycle arrest and endoplasmic reticulum (ER) stress, inhibited cell migration, reduced the % of ALDH positive cells in the population and enhanced PARP1 cleavage.Conclusion: The synergistic effect of the niraparib + F7 may result from the treatment affecting multiple genetic pathways involving cell death and reducing mesenchymal characteristics.</p