Journal: Oncology reports

Article Title: Paclitaxel enhances tumoricidal potential of TRAIL via inhibition of MAPK in resistant gastric cancer cells.

doi: 10.3892/or.2016.4666

Figure Lengend Snippet: Figure 4. PTX altered expression of death receptors and anti-apoptotic proteins. (A) DR4 and DR5 expression was assessed by western blotting in SGC-7901 and MGC-803 cells after 24-h incubation with PTX at indicated doses. (B) Cell surface expression of DR4 and DR5 was assessed by flow cytometric method in SGC-7901 and MGC-803 cells after incubation with 100 nM PTX for 24 h. The blue lines indicate staining with isotype control, black lines and red lines indicate absence and presence of PTX, respectively. (C) SGC-7901 and MGC-803 cells were treated with TRAIL (100 ng/ml) and/or PTX (1 µM) for 24 h. Anti-apoptotic proteins were analyzed by western blotting. (D) SGC-7901 and MGC-803 cells were treated with indicated doses of PTX for 24 h or 1 µM PTX for indicated time-points. Anti-apoptotic proteins were analyzed by western blotting. (E) Effect of C-IAP1 silencing on cell viability induced by TRAIL plus PTX. SGC-7901 and MGC-803 cells were transiently transfected with C-IAP1 specific siRNA for 48 h and then incubated with TRAIL (100 ng/ml) and/or PTX (10 nM) for 24 h. Cell viability was analyzed by the CCK-8 assay. **p<0.05, ***p<0.01.

Article Snippet: C-IAP1 siRNA and scrambled siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Western Blot, Incubation, Cytometry, Staining, Control, Transfection, CCK-8 Assay

Journal: Frontiers in Immunology

Article Title: Differential early response of monocyte/macrophage subsets to intra-operative corticosteroid administration in lung transplantation

doi: 10.3389/fimmu.2023.1281546

Figure Lengend Snippet: The recruited monocytic cells in lung grafts include CD16 pos and CD14 pos subsets. (A) The live CFSE pos SSC-A lo cells were selected as we reported in . The CD172A hi cells were gated and were further split into CD14 pos and CD16 pos cells. The two subsets were sorted by flow cytometry and stained with May-Grunwald Giemsa. The represented gating strategy is shown on a pig cross-circulation experiment at 10 h, depicted as a cross symbol in other figures. (B) The expression of CD163, MHC class II, and CD80/86 on the CD16 pos and CD14 pos cells of the live CFSE pos SSC-A lo CD172A hi lung cells is depicted (filled blue histogram) versus an IgG2a isotype control (black line) at 6 h and 10 h post-reperfusion. The percent of positive cells are indicated.

Article Snippet: We found that in the UT group, the percent of MHC class II + cells among the CFSE pos SSC lo CD172A hi MoCs increased from 21.8 ± 7.1% at 6 h to 38.6 ± 3% at 10 h (p = 0.008), whereas in the CST group it was only 5.3 ± 3.7% at 6 h and 3.8 ± 2.2% at 10 h (p < 0.01 between UT and CST in all cases, left panel and for representative FACS raw profiles).

Techniques: Flow Cytometry, Staining, Expressing, Control

Journal: Frontiers in Immunology

Article Title: Differential early response of monocyte/macrophage subsets to intra-operative corticosteroid administration in lung transplantation

doi: 10.3389/fimmu.2023.1281546

Figure Lengend Snippet: Representation of immune cell subsets among the CFSE pos cells at 6 h (A) and 10 h (B) in lung grafts. The gating strategy has been described in and is also shown Supplemental Figure 2 . Within live CFSE pos gated cells, the percent of PMNs, SSC-A lo CD172A hi cells (MoCs, designated CD172A hi ), CD14 pos SSC-A lo CD172A hi (designated CD14 pos ), CD16 pos SSC-A lo CD172A hi (designated CD16 pos ), SSC-A lo CD172 int cells (designated CD172A int ), CD13 pos (cDC1 dendritic cells), NKp46 pos (NK cells), CD21 pos (B cells), CD3 pos (T cells), CD3 pos CD4 pos , CD3 pos CD8 pos T cells are reported for the untreated group (UT in red) and the corticosteroid-treated (CST in black) group (5 pigs per group except for CD14 pos and CD16 pos subsets, with 3 in the UT group and 4 in the CST group). Each pig is labeled with a unique colored symbol throughout the paper. In most cases the data distribution passed the normality test and an unpaired t-test was used to identify statistically significant differences between UT and CST values; a Mann-Whitney test was used for the CD14 pos and CD16 pos subset values. No statistically significant differences were found in any case.

Article Snippet: We found that in the UT group, the percent of MHC class II + cells among the CFSE pos SSC lo CD172A hi MoCs increased from 21.8 ± 7.1% at 6 h to 38.6 ± 3% at 10 h (p = 0.008), whereas in the CST group it was only 5.3 ± 3.7% at 6 h and 3.8 ± 2.2% at 10 h (p < 0.01 between UT and CST in all cases, left panel and for representative FACS raw profiles).

Techniques: Labeling, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Differential early response of monocyte/macrophage subsets to intra-operative corticosteroid administration in lung transplantation

doi: 10.3389/fimmu.2023.1281546

Figure Lengend Snippet: Modulation of MHC class II and CD80/86 expression on lung monocytic cells and alveolar macrophages upon cross-circulation and effects of corticosteroid treatment. (A) Percent of MHC class II pos cells within live CFSE pos SSC-A lo CD172 hi cells (left panel), within live CFSE neg SSC-A lo CD172 hi cells (middle panel) and geometric mean expression of MHC class II on alveolar macrophages (all alveolar macrophages are MHC class II pos ). (B) Same as in A for the analysis of CD80/86 expression. In (A, B) , 5 pigs were monitored per group. Each pig is labeled with a unique colored symbol throughout the paper (untreated group (UT) and corticosteroid-treated (CST), UT in red, CST in black). As values passed a normality test, a paired t-test was used to identify statistically significant differences between timings, and an unpaired t-test was used to identify statistically significant differences between the UT and CST groups, with p-values reported in the table above each panel. NS stands for non-significant.

Article Snippet: We found that in the UT group, the percent of MHC class II + cells among the CFSE pos SSC lo CD172A hi MoCs increased from 21.8 ± 7.1% at 6 h to 38.6 ± 3% at 10 h (p = 0.008), whereas in the CST group it was only 5.3 ± 3.7% at 6 h and 3.8 ± 2.2% at 10 h (p < 0.01 between UT and CST in all cases, left panel and for representative FACS raw profiles).

Techniques: Expressing, Labeling

Journal: Frontiers in Immunology

Article Title: Differential early response of monocyte/macrophage subsets to intra-operative corticosteroid administration in lung transplantation

doi: 10.3389/fimmu.2023.1281546

Figure Lengend Snippet: Cytokine gene expression in the CD14 pos and CD16 pos subsets and alveolar macrophages upon cross-circulation and effects of CS treatment. Gene expression arbitrary values were calculated from RT-qPCR data normalized to a house keeping gene (RSP24) and to an internal calibrator, using the 2 -ΔΔCT method; the gene expression data were obtained from sorted CFSE pos CD172A pos CD14 pos cells, CFSE pos CD172A pos CD16 pos cells and alveolar macrophages from the untreated group (UT) and the corticosteroid-treated group (CST) at 6 h and 10 h cross-circulation (3 pigs from UT group in red, 4 pigs from the CST group in black). See the calculation method in the material and methods. As the data did not pass a normality test, a paired Wilcoxon test was used to identify statistically significant differences between timings, and a one-tailed Mann-Whitney test was used to identify statistically significant differences between the UT and CST groups, with p-values reported in the table above each panel. NS stands for non-significant.

Article Snippet: We found that in the UT group, the percent of MHC class II + cells among the CFSE pos SSC lo CD172A hi MoCs increased from 21.8 ± 7.1% at 6 h to 38.6 ± 3% at 10 h (p = 0.008), whereas in the CST group it was only 5.3 ± 3.7% at 6 h and 3.8 ± 2.2% at 10 h (p < 0.01 between UT and CST in all cases, left panel and for representative FACS raw profiles).

Techniques: Gene Expression, Quantitative RT-PCR, One-tailed Test, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Differential early response of monocyte/macrophage subsets to intra-operative corticosteroid administration in lung transplantation

doi: 10.3389/fimmu.2023.1281546

Figure Lengend Snippet: Ratio of IL10 versus inflammatory cytokine gene expression in the CD14 pos and CD16 pos subsets and in alveolar macrophages upon cross-circulation and effects of CS treatment. A ratio between the IL10 gene expression values and the different inflammatory cytokine values presented in Figure 4 was calculated for sorted CFSE pos CD172A pos CD14 pos cells, CFSE pos CD172A pos CD16 pos cells, and alveolar macrophages from the untreated group (UT) and the corticosteroid-treated group (CST) at 6 h and 10 h cross-circulation (3 pigs from UT group in red, 4 pigs from the CST group in black). As the data did not pass a normality test, a paired Wilcoxon test was used to identify statistically significant differences between timings, and a one-tailed Mann-Whitney test was used to identify statistically significant differences between the UT and CST groups, with p-values reported in the table above each panel. NS stands for non-significant.

Article Snippet: We found that in the UT group, the percent of MHC class II + cells among the CFSE pos SSC lo CD172A hi MoCs increased from 21.8 ± 7.1% at 6 h to 38.6 ± 3% at 10 h (p = 0.008), whereas in the CST group it was only 5.3 ± 3.7% at 6 h and 3.8 ± 2.2% at 10 h (p < 0.01 between UT and CST in all cases, left panel and for representative FACS raw profiles).

Techniques: Gene Expression, One-tailed Test, MANN-WHITNEY

Journal: Stroke

Article Title: Ischemic Preconditioning Reduces Neurovascular Damage After Hypoxia-Ischemia Via the Cellular Inhibitor of Apoptosis 1 in Neonatal Brain

doi: 10.1161/strokeaha.112.677617

Figure Lengend Snippet: Figure 2. A, The no ischemic preconditioning (IP) group had significantly increased cleaved caspase-8, caspase-9, caspase-3, and poly (ADP-ribose) polymerase (PARP) 24 hours post-hypoxia-ischemia (HI) compared with the IP group. B, The IP group showed significantly increased cellular inhibitor of apoptosis 1 (cIAP1) but not cIAP2, X-linked IAP (XIAP), or survivin. Data from 4 different experiments; *P<0.05.

Article Snippet: Lentivirus-mediated short hairpin RNA targeting cIAP1 mRNA (LV-sh-cIAP1; sc-29848-V; Santa Cruz) and nontarget shRNA control lentiviral particles encoding a scrambled shRNA sequence (LV-sh-scramble control; sc-108080) were used.

Techniques:

Journal: Stroke

Article Title: Ischemic Preconditioning Reduces Neurovascular Damage After Hypoxia-Ischemia Via the Cellular Inhibitor of Apoptosis 1 in Neonatal Brain

doi: 10.1161/strokeaha.112.677617

Figure Lengend Snippet: Figure 3. A, Immunohistochemistry showed that cellular inhibitor of apoptosis 1 (cIAP1) was markedly decreased in the group with no ischemic preconditioning (IP) but increased in the IP group 24-hour post-hypoxia-ischemia (HI). In the IP group, cIAP1 was mainly expressed in the vascular (arrows) and nonvascular cells. Scale bar, 100 μm. B, Immunofluorescence confirmed that cIAP1 expressed mainly in the rat endothelial cell antigen-1-positive endothelial cells and NeuN-positive neurons, but not in astrocytes (glial fibrillary acidic protein [GFAP]), in IP group. C, Compared with control, cIAP1 was significantly upregulated in IP group pretreated with control small interfering RNA (siRNA) but not in that pretreated with cIAP1 siRNA. n=4 per group. D, In IP group, the pups pretreated with cIAP1 siRNA had significantly more brain damage than those with control siRNA. n=7 per group. Scale bar, 20 μm. *P<0.05; **P<0.01; #P<0.001.

Article Snippet: Lentivirus-mediated short hairpin RNA targeting cIAP1 mRNA (LV-sh-cIAP1; sc-29848-V; Santa Cruz) and nontarget shRNA control lentiviral particles encoding a scrambled shRNA sequence (LV-sh-scramble control; sc-108080) were used.

Techniques: Immunohistochemistry, Immunofluorescence, Control, Small Interfering RNA

Journal: Stroke

Article Title: Ischemic Preconditioning Reduces Neurovascular Damage After Hypoxia-Ischemia Via the Cellular Inhibitor of Apoptosis 1 in Neonatal Brain

doi: 10.1161/strokeaha.112.677617

Figure Lengend Snippet: Figure 4. SH-SY5Y neurons. A, Left, Compared with controls, cytotoxicity increased progressively when oxygen- glucose deprivation (OGD) duration increased to 15 hours (left). Right, Eight- hour OGD preconditioning before 15-hour OGD significantly decreased cytotoxicity. B, Preconditioned cells had significantly increased cellular inhibitor of apoptosis 1 (cIAP1) at 1, 6, and 24 hours post-OGD than nonpreconditioned cells. C, Lentivi rus-mediated short hairpin RNA targeting cIAP1 mRNA (LV-sh-cIAP1) cells had 25% of the cIAP1 levels of LV-sh-scram ble cells. D, Post-OGD, preconditioned LV-sh-scramble cells were significantly less cytotoxic than nonpreconditioned LV-sh-scramble cells, and preconditioned LV-sh-cIAP1 cells had significantly more cytotoxicity than preconditioned LV- sh-scramble cells. Data were from 4 different experiments. *P<0.05; #P<0.001.

Article Snippet: Lentivirus-mediated short hairpin RNA targeting cIAP1 mRNA (LV-sh-cIAP1; sc-29848-V; Santa Cruz) and nontarget shRNA control lentiviral particles encoding a scrambled shRNA sequence (LV-sh-scramble control; sc-108080) were used.

Techniques: shRNA

Journal: Stroke

Article Title: Ischemic Preconditioning Reduces Neurovascular Damage After Hypoxia-Ischemia Via the Cellular Inhibitor of Apoptosis 1 in Neonatal Brain

doi: 10.1161/strokeaha.112.677617

Figure Lengend Snippet: Figure 5. A, Left, Cytotoxicity increased progressively in human microvascular endothelial cell-1 (HMEC-1) endothelial cells when oxygen–glucose deprivation (OGD) duration increased up to 15 hours. Right, Seven-hour OGD precondition ing before 15-hour OGD significantly decreased cytotoxicity in HMEC-1 cells. B, Preconditioned cells had significantly more cellular inhibitor of apoptosis 1 (cIAP1) than nonpreconditioned cells at 24 and 48 hours post-OGD. C, Lentivirus- mediated short hairpin RNA targeting cIAP1 mRNA (LV-sh-cIAP1) cells had 45% of cIAP1 levels of LV-sh-scramble cells. D, Post-OGD, preconditioned LV- sh-cIAP1 cells had significantly more cytotoxicity than preconditioned LV- sh-scramble cells. Data from 4 different experiments. *P<0.05; #P<0.001.

Article Snippet: Lentivirus-mediated short hairpin RNA targeting cIAP1 mRNA (LV-sh-cIAP1; sc-29848-V; Santa Cruz) and nontarget shRNA control lentiviral particles encoding a scrambled shRNA sequence (LV-sh-scramble control; sc-108080) were used.

Techniques: shRNA

Journal: Stroke

Article Title: Ischemic Preconditioning Reduces Neurovascular Damage After Hypoxia-Ischemia Via the Cellular Inhibitor of Apoptosis 1 in Neonatal Brain

doi: 10.1161/strokeaha.112.677617

Figure Lengend Snippet: Figure 6. A, Lentiviruses encoding cel lular inhibitor of apoptosis 1 (LV-cIAP1) SH-SY5Y neurons had 30% increases in cIAP1 compared with LV-control neurons. B, After oxygen–glucose depri vation (OGD), LV-cIAP1 neurons had significantly decreased cytotoxicity than LV-control neurons. C, LV-cIAP1 human microvascular endothelial cells-1 (HMEC-1) showed 70% increases in cIAP1 compared with LV-control cells. D, Post-OGD, LV-cIAP1 cells had sig nificantly decreased cytotoxicity than LV-control cells. Data from 4 different experiments. **P<0.01; #P<0.001.

Article Snippet: Lentivirus-mediated short hairpin RNA targeting cIAP1 mRNA (LV-sh-cIAP1; sc-29848-V; Santa Cruz) and nontarget shRNA control lentiviral particles encoding a scrambled shRNA sequence (LV-sh-scramble control; sc-108080) were used.

Techniques: Control