Journal: Oncology reports

Article Title: miRNA-regulated expression of oncogenes and tumor suppressor genes in the cisplatin-inhibited growth of K562 cells.

doi: 10.3892/or_00000813

Figure Lengend Snippet: Figure 2. Changes in mRNA expression of BCL2, E2F1, E2F3, RB1 and P53 in K562 cells after cisplatin treatment. BCL2, E2F1, E2F3, RB1 and P53 were detected by (A) RT-PCR, (B) real-time PCR and (C) ELISA.

Article Snippet: K562 cells were lysed [lysis buffer: 0.15 M NaCl, 5 mM EDTA (pH 8.0), 1% Triton X-100, 10 mM Tris-Cl (pH 7.4), 100 mM PMSF and 5 M DTT) and incubated in a 96-well plate, followed by the addition of goat anti-human antibodies against BCL2 (1:400), E2F1, E2F3 (1:400, 1:400), RB1 (1:500) and P53 (1:300, 1:400) (all from Immunoleader, Boster) according to the manufacturer's instructions.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Oncology reports

Article Title: miRNA-regulated expression of oncogenes and tumor suppressor genes in the cisplatin-inhibited growth of K562 cells.

doi: 10.3892/or_00000813

Figure Lengend Snippet: Figure 5. Expression of E2F1, E2F3, BCL2, RB1 and P53 genes regulated by miRNAs. Detection of the expression of (A) E2F1, (B) E2F3, (C) BCL2, (D) RB1 and (E) P53. *Significant difference (p<0.05).

Article Snippet: K562 cells were lysed [lysis buffer: 0.15 M NaCl, 5 mM EDTA (pH 8.0), 1% Triton X-100, 10 mM Tris-Cl (pH 7.4), 100 mM PMSF and 5 M DTT) and incubated in a 96-well plate, followed by the addition of goat anti-human antibodies against BCL2 (1:400), E2F1, E2F3 (1:400, 1:400), RB1 (1:500) and P53 (1:300, 1:400) (all from Immunoleader, Boster) according to the manufacturer's instructions.

Techniques: Expressing

Journal: Oncology reports

Article Title: miRNA-regulated expression of oncogenes and tumor suppressor genes in the cisplatin-inhibited growth of K562 cells.

doi: 10.3892/or_00000813

Figure Lengend Snippet: Figure 6. Correlative expression of miRNA and tumor suppressor genes with ASO. Correlative expression of (A) RB1 and miR-106 and (B) P53 and miR-150 and their downstream factors after ASO treatment. *Significant difference (p<0.05).

Article Snippet: K562 cells were lysed [lysis buffer: 0.15 M NaCl, 5 mM EDTA (pH 8.0), 1% Triton X-100, 10 mM Tris-Cl (pH 7.4), 100 mM PMSF and 5 M DTT) and incubated in a 96-well plate, followed by the addition of goat anti-human antibodies against BCL2 (1:400), E2F1, E2F3 (1:400, 1:400), RB1 (1:500) and P53 (1:300, 1:400) (all from Immunoleader, Boster) according to the manufacturer's instructions.

Techniques: Expressing

Journal: Oncology reports

Article Title: miRNA-regulated expression of oncogenes and tumor suppressor genes in the cisplatin-inhibited growth of K562 cells.

doi: 10.3892/or_00000813

Figure Lengend Snippet: Figure 7. miRNA expression downstream of RB1 and P53 was detected by real-time PCR. (A) Expression of miR-34a-c after treatment with anti-106. (B) Expression of miR-34a-c and miR-16 after treatment with anti-150. *Significant difference (p<0.05).

Article Snippet: K562 cells were lysed [lysis buffer: 0.15 M NaCl, 5 mM EDTA (pH 8.0), 1% Triton X-100, 10 mM Tris-Cl (pH 7.4), 100 mM PMSF and 5 M DTT) and incubated in a 96-well plate, followed by the addition of goat anti-human antibodies against BCL2 (1:400), E2F1, E2F3 (1:400, 1:400), RB1 (1:500) and P53 (1:300, 1:400) (all from Immunoleader, Boster) according to the manufacturer's instructions.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: International journal of biological sciences

Article Title: Integrative analysis of multi-omics data reveals inhibition of RB1 signaling promotes apatinib resistance of hepatocellular carcinoma.

doi: 10.7150/ijbs.83862

Figure Lengend Snippet: Figure 4. Changes in chromatin accessibility are consistent with global transcriptome changes. A. Genome tracks showing a comparison of ATAC-seq profiles of RB1 in apatinib-resistant and non-resistant HepG2 cells. B. The mRNA expression levels of RB1 in apatinib-resistant versus untreated HepG2 cells, statistical analysis was performed by Student t-test. C. Boxplots of chromatin accessibility values for high expression genes and low expression genes in apatinib-resistant cells.

Article Snippet: After overnight incubation with primary antibody (anti - RB1 and CDKN2B antibody; Boster Bio, Pleasanton, USA) by 1:200 at 4 °C, sections were incubated for 15 minutes at room temperature with horseradish peroxidaselabeled polymer conjugated with secondary antibody (MaxVision Kits) and incubated for 1 minute with diaminobenzidine.

Techniques: Comparison, Expressing

Journal: International journal of biological sciences

Article Title: Integrative analysis of multi-omics data reveals inhibition of RB1 signaling promotes apatinib resistance of hepatocellular carcinoma.

doi: 10.7150/ijbs.83862

Figure Lengend Snippet: Figure 6. Expression status of RB1 pathway targets in HCC cells and in clinical samples; CDK2-IN-73 and Palbociclib can relieve apatinib resistance. A. qPCR analysis showed that the expression levels of RB1, PCNA, KIAA0101 and MCM7 in HepG2-resistant cells were significantly reduced, and the expression levels of CDKN2B andCDKN2AIP were increased. B. Western blot detection of the above proteins is consistent with qPCR detection results. C. Analysis of mRNA expression of five PD patients showed high expression of CDKN2B and low expression of RB1, PCNA, and MCM7. D. Western blot analysis showed that PD patients had high expression of CDKN2B but low expression of RB1, PCNA, and MCM7. E. Representative photographs of staining of RB1 protein in a pair of HCC patients tissues in PD and PR (20X). F. Representative photographs of staining of CDKN2B protein in a pair of HCC patients tissues in PD and PR (20X). G. The proliferation inhibition curves of resistant HepG2 cells and HepG2 cells treated with CDK2-IN-73. H. Western blot analysis showed that the expression of CDKN2B in HepG2resistant cells was not significantly different, and the expression levels of RB1, PCNA and MCM7 were upregulated. I. Proliferation inhibition curves of HepG2 resistant cells and HepG2 cells treated with Palbociclib. J. Western blot analysis results showed that the expression levels of RB1, PCNA, and MCM7 were upregulated. The bars represent mean ± standard deviation; ∗P < 0.05.

Article Snippet: After overnight incubation with primary antibody (anti - RB1 and CDKN2B antibody; Boster Bio, Pleasanton, USA) by 1:200 at 4 °C, sections were incubated for 15 minutes at room temperature with horseradish peroxidaselabeled polymer conjugated with secondary antibody (MaxVision Kits) and incubated for 1 minute with diaminobenzidine.

Techniques: Expressing, Western Blot, Staining, Inhibition, Standard Deviation

Journal: Aging (Albany NY)

Article Title: Molecular characterization of long-term survivors of hepatocellular carcinoma

doi: 10.18632/aging.202615

Figure Lengend Snippet: Abemaciclib substantially inhibited the proliferation of HCC cells in vitro . ( A ) The protein levels of retinoblastoma (p-S780) and retinoblastoma in Hep3B and Huh7 cells. ( B , C ) Clone formation assays were used to detect the effects of abemaciclib on the proliferation of Hep3B and Huh7 cells. ( D ) A CCK-8 assay was used to analyze cell proliferation in control and abemaciclib-treated Hep3B and Huh7 cells. ( E ) An Edu assay was used to analyze cell proliferation in control and abemaciclib-treated Hep3B and Huh7 cells. Bar= 100μm; For ( C , D ), * was compared with Control (**, P < 0.01; ***, P < 0.001).

Article Snippet: The membranes were incubated overnight at 4° C with primary antibodies for retinoblastoma (phospho S780) (ab47763, Abcam, Cambridge, UK), retinoblastoma (ab181616) and GAPDH (BM1623, Boster, Wuhan, China), and then were incubated with the appropriate horseradish peroxidase-conjugated secondary antibodies for 2 hours.

Techniques: In Vitro, CCK-8 Assay, Control, EdU Assay

Journal: Oncotarget

Article Title: PDGFRα depletion attenuates glioblastoma stem cells features by modulation of STAT3, RB1 and multiple oncogenic signals

doi: 10.18632/oncotarget.10132

Figure Lengend Snippet: A. Time course experiment of the effects of PDGF-AA on immediate early genes activation in c-CSC2 vs p-CSC2 after 2 days of growth factors starvation. p-CSC2 express Egr1 and Y705-phosphorylated Stat3 after 5 minutes, on the other hand, hypo-Rb1 is significantly downmodulated in p-CSC2 after 10 minutes of treatment. Instead, the levels of T638/641-PKCα/βII remain stable at all time points. B. PDGF-AA applied alone or with CR for 24 hours on p-CSC2. Stat3 Y705 phosphorylation levels are impaired by CR treatment, instead hypo-Rb1 and total Rb1 protein expression are not affected after 24 hours by PDGFR inhibition. Cyclin D1 protein levels are not modulated in the same conditions. β-actin is used as control of equal proteins loading. C. Time course experiment to evaluate the effects of PDGF-AA alone or combined with CR on the regulation of hypo-Rb1 in p-CSC2. CR upregulates hypo-Rb1 expression respect to PDGF-AA alone.

Article Snippet: Filters were immunoblotted using the following primary antibodies: rabbit anti-EGFR, rabbit anti-Y1068-EGFR, goat anti-PDGFRα, rabbit anti-T202/Y204-ERK1/2 and anti-ERK1/2, rabbit anti-Y705-STAT3 and rabbit anti-STAT3, rabbit anti-T638/641-PKCα/βII, rabbit anti-PCKα, rabbit anti-VEGFR2 (all purchased from Cell Signaling, MA-USA), mouse anti-underphosphorylated Rb1 (BD Pharmigen), Rabbit anti-Rb1 (Cell Signaling), mouse anti-BCL-2 (Dako), mouse anti-Nestin (Millipore), mouse anti-GFAP (Covance), rabbit anti-NG2 (Chemicon), mouse anti-MAP2a/b and rabbit anti-OCT4 (Millipore), rabbit anti-BCL-x/L, rabbit anti-CycD1, rabbit anti-SOX2, rabbit anti-BMI1 and rabbit anti-EGR1, rabbit anti-CD31, goat anti-VE-Cadherin (Santa Cruz-USA), mouse anti-β-actin (SIGMA).

Techniques: Activation Assay, Expressing, Inhibition

Journal: Oncotarget

Article Title: PDGFRα depletion attenuates glioblastoma stem cells features by modulation of STAT3, RB1 and multiple oncogenic signals

doi: 10.18632/oncotarget.10132

Figure Lengend Snippet: A. Western blot analysis reveals that depletion of PDGFRα in p-CSC2 induces the downmodulation of Stat3 Y705 phosphorylation in both proliferating cell clones, instead their levels raised up in differentiation conditions. The hypo-Rb1 levels result overexpressed in clone 3 in all conditions. Instead the clone 1, shows an increase of hypo-Rb1 at 2 and 4 day of differentiation with respect to pLKO.1 cells. In parallel, Bcl-2 and Bcl-xL proteins and Erk1/2 pathway were upregulated in both cell clones in comparison with pLKO.1 cells in differentiaton conditions. T638/641 phosphorylated-PKCα/βII isoforms expression levels are slightly induced in proliferating PDGFRα-depleted cell clones, but negatively modulated in differentiated ones than pLKO.1 cells. B. The downmodulation of multipotent protein markers (Nestin, Sox2, Bmi1), except for Oct4, is accompanied with early induction of the neuronal marker MAP2a/b in proliferating PDGFRα-depleted cell clones vs pLKO.1. Similar results were observed during cell differentiation, except for MAP2a/b that increases in pLKO.1 with respect to PDGFRα-depleted cells.

Article Snippet: Filters were immunoblotted using the following primary antibodies: rabbit anti-EGFR, rabbit anti-Y1068-EGFR, goat anti-PDGFRα, rabbit anti-T202/Y204-ERK1/2 and anti-ERK1/2, rabbit anti-Y705-STAT3 and rabbit anti-STAT3, rabbit anti-T638/641-PKCα/βII, rabbit anti-PCKα, rabbit anti-VEGFR2 (all purchased from Cell Signaling, MA-USA), mouse anti-underphosphorylated Rb1 (BD Pharmigen), Rabbit anti-Rb1 (Cell Signaling), mouse anti-BCL-2 (Dako), mouse anti-Nestin (Millipore), mouse anti-GFAP (Covance), rabbit anti-NG2 (Chemicon), mouse anti-MAP2a/b and rabbit anti-OCT4 (Millipore), rabbit anti-BCL-x/L, rabbit anti-CycD1, rabbit anti-SOX2, rabbit anti-BMI1 and rabbit anti-EGR1, rabbit anti-CD31, goat anti-VE-Cadherin (Santa Cruz-USA), mouse anti-β-actin (SIGMA).

Techniques: Western Blot, Clone Assay, Expressing, Marker, Cell Differentiation

Journal: Oncotarget

Article Title: PDGFRα depletion attenuates glioblastoma stem cells features by modulation of STAT3, RB1 and multiple oncogenic signals

doi: 10.18632/oncotarget.10132

Figure Lengend Snippet: RT-qPCR analysis shows a significant modulation of gene expression of cell-cell (CDH1, CDH5) and cell-substrate (FN1) molecules interactions, the stem cell marker SOX2 and oncogenic signaling molecules (PRKCA, PRKCB1). Progressive upregulation of the tumor suppressor RB1 and PRKCB1 isoform during cell differentiation is shown. Results represent the mean of two independent experiments performed in triplicate. Fold change (Fc).

Article Snippet: Filters were immunoblotted using the following primary antibodies: rabbit anti-EGFR, rabbit anti-Y1068-EGFR, goat anti-PDGFRα, rabbit anti-T202/Y204-ERK1/2 and anti-ERK1/2, rabbit anti-Y705-STAT3 and rabbit anti-STAT3, rabbit anti-T638/641-PKCα/βII, rabbit anti-PCKα, rabbit anti-VEGFR2 (all purchased from Cell Signaling, MA-USA), mouse anti-underphosphorylated Rb1 (BD Pharmigen), Rabbit anti-Rb1 (Cell Signaling), mouse anti-BCL-2 (Dako), mouse anti-Nestin (Millipore), mouse anti-GFAP (Covance), rabbit anti-NG2 (Chemicon), mouse anti-MAP2a/b and rabbit anti-OCT4 (Millipore), rabbit anti-BCL-x/L, rabbit anti-CycD1, rabbit anti-SOX2, rabbit anti-BMI1 and rabbit anti-EGR1, rabbit anti-CD31, goat anti-VE-Cadherin (Santa Cruz-USA), mouse anti-β-actin (SIGMA).

Techniques: Quantitative RT-PCR, Expressing, Marker, Cell Differentiation

Journal: Reproductive Medicine and Biology

Article Title: Upregulated serum granulysin levels in women with antiphospholipid antibody‐associated recurrent miscarriage are downregulated by heparin treatment

doi: 10.1002/rmb2.12460

Figure Lengend Snippet: Serum granulysin concentration in women with or without antiphospholipid antibodies

Article Snippet: Briefly, microtiter plates (Nunc) were coated with 5 mg/ml anti‐granulysin monoclonal (m)Ab RB1(Mouse IgG1κ) (MBL International Corporation) in 100 mM carbonate buffer and maintained overnight at 4°C.

Techniques: Concentration Assay

Journal: Reproductive Medicine and Biology

Article Title: Upregulated serum granulysin levels in women with antiphospholipid antibody‐associated recurrent miscarriage are downregulated by heparin treatment

doi: 10.1002/rmb2.12460

Figure Lengend Snippet: Changes in serum granulysin levels in patients with RPL treated with heparin and aspirin. Serum granulysin concentration was assayed using ELISA. (a) Serum granulysin levels of patients treated with heparin plus aspirin combination therapy were measured before and 1 week after the initiation of heparin treatment ( n = 32). (b) Serum granulysin levels of patients treated with aspirin alone ( n = 30). The gestational age of sampling is the same as in the heparin plus aspirin combination therapy group. *** p < 0.001: paired t test

Article Snippet: Briefly, microtiter plates (Nunc) were coated with 5 mg/ml anti‐granulysin monoclonal (m)Ab RB1(Mouse IgG1κ) (MBL International Corporation) in 100 mM carbonate buffer and maintained overnight at 4°C.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Sampling

Journal: Reproductive Medicine and Biology

Article Title: Upregulated serum granulysin levels in women with antiphospholipid antibody‐associated recurrent miscarriage are downregulated by heparin treatment

doi: 10.1002/rmb2.12460

Figure Lengend Snippet: Changes in serum granulysin levels in patients with PE‐positive and PE‐negative RPL. Serum granulysin concentration was assayed using ELISA. (a, b) Serum granulysin levels of PE‐positive (a) and PE‐negative (b) patients treated with heparin plus aspirin were measured before and 1 week after the initiation of heparin treatment ( n = 19 and 13, respectively). (The gestational age of sampling is the same as in the heparin plus aspirin combination therapy group.) ** p < 0.01: paired t test

Article Snippet: Briefly, microtiter plates (Nunc) were coated with 5 mg/ml anti‐granulysin monoclonal (m)Ab RB1(Mouse IgG1κ) (MBL International Corporation) in 100 mM carbonate buffer and maintained overnight at 4°C.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Sampling

Journal: Reproductive Medicine and Biology

Article Title: Upregulated serum granulysin levels in women with antiphospholipid antibody‐associated recurrent miscarriage are downregulated by heparin treatment

doi: 10.1002/rmb2.12460

Figure Lengend Snippet: Pregnancy outcome stratified by granulysin levels after heparin plus aspirin treatment. ** p < 0.05: Fisher's exact probability test

Article Snippet: Briefly, microtiter plates (Nunc) were coated with 5 mg/ml anti‐granulysin monoclonal (m)Ab RB1(Mouse IgG1κ) (MBL International Corporation) in 100 mM carbonate buffer and maintained overnight at 4°C.

Techniques:

Journal: PLoS ONE

Article Title: Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin

doi: 10.1371/journal.pone.0156321

Figure Lengend Snippet: Antimicrobial activities of 15 kDa granulysin on Gram-positive (A) and Gram-negative (B) bacteria. Microbes (5–10×10 4 cfu) were incubated with 15 kDa granulysin at 37°C in 10 mM sodium phosphate, pH 7.5, for 3 hr and plated on agar plates for the determination of remaining cfu. Comparison on the antimicrobial activity against P . aeruginosa among 15 kDa, 9 kDa granulysin and a commercial 15 kDa granulysin (R&D) were also employed (C). S . a ., Staphylococcus aureus ; E . f ., Enterococcus faecalis ; L . i ., Listeria innocua ; P . a ., Pseudomonas aeruginosa ; Y . e ., Yersinia enterocolitica ; S . m ., Serratia marcescens ; E . c ., Escherichia coli ; S . t ., Salmonella typhimurium ; K . p ., Klebsiella pneumonia .

Article Snippet: The cells were fixed with 4% paraformaldehyde, permeabilized by 0.5% Triton X-100, and incubated with anti-granulysin antibody RF10 (MBL, Chicago, USA) (1:1000) and Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (Molecular Probes, OR, USA) (1:1000) for the localization of granulysin.

Techniques: Incubation, Activity Assay

Journal: PLoS ONE

Article Title: Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin

doi: 10.1371/journal.pone.0156321

Figure Lengend Snippet: Approximately 1×10 7 microbes were incubated with 2 μM 15 kDa granulysin for 0 min (A), 60 min (B) and 180 min (C), respectively. Immunocytochemistry was performed using RF10 mAb for granulysin staining and DAPI for DNA staining. Images were taken using confocal laser-scanning microscopy (CLSM).

Article Snippet: The cells were fixed with 4% paraformaldehyde, permeabilized by 0.5% Triton X-100, and incubated with anti-granulysin antibody RF10 (MBL, Chicago, USA) (1:1000) and Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (Molecular Probes, OR, USA) (1:1000) for the localization of granulysin.

Techniques: Incubation, Immunocytochemistry, Staining, Confocal Laser Scanning Microscopy

Journal: PLoS ONE

Article Title: Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin

doi: 10.1371/journal.pone.0156321

Figure Lengend Snippet: Approximately 1×10 7 microbes were treated with 2 μM 15 kDa granulysin for 0 min (A-C) and 180 min (D-F) and observed by transmission electron microscopy (TEM).

Article Snippet: The cells were fixed with 4% paraformaldehyde, permeabilized by 0.5% Triton X-100, and incubated with anti-granulysin antibody RF10 (MBL, Chicago, USA) (1:1000) and Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (Molecular Probes, OR, USA) (1:1000) for the localization of granulysin.

Techniques: Transmission Assay, Electron Microscopy

Journal: PLoS ONE

Article Title: Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin

doi: 10.1371/journal.pone.0156321

Figure Lengend Snippet: Membrane potential of SMAP-29- (A) and 15 kDa granulysin- (B) treated microbes were determined in the presence of DiSC 3 (5). Fluorescence intensity was monitored at an excitation wavelength of 622 nm and an emission wavelength of 670 nm. Data plotted are representative average values of three independent trials. a.u., absorbance unit. Membrane potential of treated microbes were also measured in the presence of DiOC 2 (3) by flow cytometry (C). Red/green fluorescence ratio was calculated using population mean fluorescence intensities for bacteria in the presence of SMAP-29 or 15kDa granulysin. Membrane permeability of microbes was monitored in the presence of SYTOX ™ Green at 485 nm and 520 nm for excitation and emission wavelength, respectively (D). Data plotted are normalized with values of untreated sample and representative average values of three independent trials. RFU, relative fluorescence unit.

Article Snippet: The cells were fixed with 4% paraformaldehyde, permeabilized by 0.5% Triton X-100, and incubated with anti-granulysin antibody RF10 (MBL, Chicago, USA) (1:1000) and Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (Molecular Probes, OR, USA) (1:1000) for the localization of granulysin.

Techniques: Fluorescence, Flow Cytometry, Permeability

Journal: PLoS ONE

Article Title: Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin

doi: 10.1371/journal.pone.0156321

Figure Lengend Snippet: The antimicrobial activities of 2 μM 15 kDa granulysin were determined in the presence of NaCl (A) and MgCl 2 (B) at the concentrations indicated. Microbes were incubated with granulysin at different pH values and the remaining colony-forming units were determined (C). SYTOX ™ Green was added to monitor the change in bacterial membrane permeability in the presence of 1 μM granulysin at pH 5.4 and pH 7.4, respectively (D). Fluorescence emission spectra of 8-Anilino-1-naphthalenesulfonic acid (ANS) was excited at 370 nm and measured between 400 and 600 nm in the presence of 1 μM granulysin at pH 7.4 (E) and pH 5.4 (F). Arrow indicates the blue shift (470 nm) of emission spectra of ANS-granulysin hydrophobic complex. Data plotted are normalized by the fluorescence intensity of ANS alone at pH 7.4 or pH 5.4, respectively. NFI, normalized fluorescence intensity. Circular dichroism spectrum of 20 μM 15 kDa granulysin in 20 mM HEPES, 50 mM NaCl, pH 7.4 or pH 5.4 (G). M.E., molar ellipticity.

Article Snippet: The cells were fixed with 4% paraformaldehyde, permeabilized by 0.5% Triton X-100, and incubated with anti-granulysin antibody RF10 (MBL, Chicago, USA) (1:1000) and Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (Molecular Probes, OR, USA) (1:1000) for the localization of granulysin.

Techniques: Incubation, Permeability, Fluorescence

Journal: PLoS ONE

Article Title: Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin

doi: 10.1371/journal.pone.0156321

Figure Lengend Snippet: Microbes were grown in M63 minimal media in the presence of SMAP-29 and 15 kDa granulysin at 37°C for 24 hr and quantified by crystal violet staining and measured at the absorbance of 600 nm. Error bars represent the standard errors of the means. The asterisks indicate samples that are significantly different from control. (Student’s t-tests; p≤0.01).

Article Snippet: The cells were fixed with 4% paraformaldehyde, permeabilized by 0.5% Triton X-100, and incubated with anti-granulysin antibody RF10 (MBL, Chicago, USA) (1:1000) and Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (Molecular Probes, OR, USA) (1:1000) for the localization of granulysin.

Techniques: Staining

Journal: PLoS ONE

Article Title: Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin

doi: 10.1371/journal.pone.0156321

Figure Lengend Snippet: Microbes were visualized by staining with SYTO-9 (green fluorescence for living bacteria) and propidium iodide (red fluorescence for dead bacteria) in the presence of 0 (A), 0.25 μM (B), and 4 μM granulysin (C) at 37°C for 24 hr. Images were obtained by confocal laser scanning microscope (CLSM).

Article Snippet: The cells were fixed with 4% paraformaldehyde, permeabilized by 0.5% Triton X-100, and incubated with anti-granulysin antibody RF10 (MBL, Chicago, USA) (1:1000) and Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (Molecular Probes, OR, USA) (1:1000) for the localization of granulysin.

Techniques: Staining, Fluorescence, Laser-Scanning Microscopy