Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes-8

<p><b>Copyright information:</b></p><p>Taken from "Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes"</p><p>http://www.biomedcentral.com/1471-2172/8/9</p><p>BMC Immunology 2007;8():9-9.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1914365.</p><p></p>les were taken for RT-PCR analysis. Representative gel image shows transcripts present in LAK cells over the stimulation period (A). The same transcripts were found in different subpopulations (not shown). Graph (B) represents results of semiquantitative analysis of the transcripts normalized to actin (gel image, lines A) in LAK cells from three different donors. Each symbol represents one donor and bars correspond to the mean value. Graph (C) shows the same analysis in different subpopulations where one representative donor is shown

Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes-4

<p><b>Copyright information:</b></p><p>Taken from "Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes"</p><p>http://www.biomedcentral.com/1471-2172/8/9</p><p>BMC Immunology 2007;8():9-9.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1914365.</p><p></p>r 10 μm

LAK cells (A) or different subpopulations (B) were stained for granulysin and CD surface markers

<p><b>Copyright information:</b></p><p>Taken from "Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes"</p><p>http://www.biomedcentral.com/1471-2172/8/9</p><p>BMC Immunology 2007;8():9-9.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1914365.</p><p></p> Cells were analysed using confocal laser scanning microscope by counting 500 cells pro sample. Graph (C) represents phenotype analysis of LAK cells. Four different donors were analysed and each symbol represents one donor. Bars indicate mean values. (* p < 0.05; * * p < 0.005; * * * p < 0.0005

Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes-0

<p><b>Copyright information:</b></p><p>Taken from "Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes"</p><p>http://www.biomedcentral.com/1471-2172/8/9</p><p>BMC Immunology 2007;8():9-9.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1914365.</p><p></p>d in LAK or antigen specific T-cells. Black boxes indicate coding and gray boxes noncoding exon parts. Exons are marked with roman numbers. Dotted lines correspond to intron regions. Arabic numbers 1 to 7 stay for primers numbered in the same way as in Table 1

Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes-2

<p><b>Copyright information:</b></p><p>Taken from "Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes"</p><p>http://www.biomedcentral.com/1471-2172/8/9</p><p>BMC Immunology 2007;8():9-9.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1914365.</p><p></p>ds. Samples were collected before stimulation (day 0), before restimulation (day 6) and 6 days after restimulation (day 12). PCR was performed and analysed by gel electrophoresis. Values are results from semiquantitative analysis of transcripts normalised to actin from the same cDNA. Each symbol represents one donor

Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes-7

<p><b>Copyright information:</b></p><p>Taken from "Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes"</p><p>http://www.biomedcentral.com/1471-2172/8/9</p><p>BMC Immunology 2007;8():9-9.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1914365.</p><p></p>d in LAK or antigen specific T-cells. Black boxes indicate coding and gray boxes noncoding exon parts. Exons are marked with roman numbers. Dotted lines correspond to intron regions. Arabic numbers 1 to 7 stay for primers numbered in the same way as in Table 1

Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes-6

<p><b>Copyright information:</b></p><p>Taken from "Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes"</p><p>http://www.biomedcentral.com/1471-2172/8/9</p><p>BMC Immunology 2007;8():9-9.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1914365.</p><p></p>f cells and separation of proteins by SDS-PAGE. Proteins were blotted and detected using anti-granulysin Ab (1:1000). The blot was stripped and probed for the β-actin

Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes-5

<p><b>Copyright information:</b></p><p>Taken from "Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes"</p><p>http://www.biomedcentral.com/1471-2172/8/9</p><p>BMC Immunology 2007;8():9-9.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1914365.</p><p></p>. Cells were analysed with a confocal laser scanning microscope counting 300 cells. Graph (A) shows granulysin expression in antigen specific T-cells as a whole population; (B) represent phenotype of specific T cells; (C) phenotype of L. specific T cells graph (D) represent granulysin expression in different subsets in specific T cells and graph (E) in specific T cells. In (B), (C), (D), and (E) every donor is indicated with one symbol. Bars are mean values. (*** p < 0.0005

Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells-6

<p><b>Copyright information:</b></p><p>Taken from "Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells"</p><p>http://www.biomedcentral.com/1471-2172/8/14</p><p>BMC Immunology 2007;8():14-14.</p><p>Published online 16 Aug 2007</p><p>PMCID:PMC1976101.</p><p></p>ith 2.5 μM granulysin and 2.5 kU/ml perforin (B) for 3 hours at 37°C. After fixation, the cells were stained with an anti-His Ab. Nuclear and bacterial DNA was labeled with DAPI (green), and granulysin is depicted in red, colocalization of the DAPI and granulysin was calculated and visualized using Imaris software (lower row), bar = 8 μm. At indicated time points colocalization of bacterial DNA with the granulysin staining was quantified in four independent experiments (C) Mean values and SD of colocalization in all evaluated cells per time point of four independent experiments are presented

Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells-4

<p><b>Copyright information:</b></p><p>Taken from "Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells"</p><p>http://www.biomedcentral.com/1471-2172/8/14</p><p>BMC Immunology 2007;8():14-14.</p><p>Published online 16 Aug 2007</p><p>PMCID:PMC1976101.</p><p></p>anulysin and 2.5 kU/ml perforin (B) at 37°C, or at 4°C (C and D). As a control, cells were treated with 2.5 μM actinand 2.5 kU/ml perforin (E). After the incubation, the cells were fixed and stained with the anti-His Ab for CLSM. Representative phase contrast (right panels) and immunofluorescence images (left panels) are shown, bar = 8 μm. -challenged DC were treated for 45 minutes with 2.5 μM granulysin and indicated concentrations of perforin at 4°C (E) or 37°C (F). After the incubation the DC were washed with PBS or with 10 mM arginine in PBS before lysis with 0.5% Triton-X-100 in PBS. The level of bound or internalized granulysin was analysed by Western blotting. Granulysin was detected with the anti-His mAb. As a reference, cellular actin was detected using an anti-β-actin mAb. Relative intensity of the bands was determined using Image J software. Mean values and SD of four independent experiments are presented