Journal: American Journal of Translational Research

Article Title: LncRNA AL645608.3 mediates malignant progression of acute myeloid leukemia

doi:

Figure Lengend Snippet: Primer information for RT-qPCR

Article Snippet: The transferred membranes were blocked for 1 h. After discarding the sealing liquid, the diluted primary antibodies against IRF6 (#sc-37704; 1:500; Santa, Texas, USA), CBL (#25818-1-AP; 1:1000; Proteintech, Wuhan, China), IFNB1 (#ab275880; 1:500; Abcam, Massachusetts, USA), and GAPDH (#ab181602; 1:10000; Abcam) were added and incubated overnight at 4°C.

Techniques: Sequencing

Journal: American Journal of Translational Research

Article Title: LncRNA AL645608.3 mediates malignant progression of acute myeloid leukemia

doi:

Figure Lengend Snippet: Effects of AL645608.3 on the mRNA expression of IRF6, CBL and IFNB1 in AML cells. A-C. The mRNA expression of IRF6, CBL and IFNB1 in THP-1 cells infected with sh-AL645608.3 lentiviruses. D-F. The mRNA expression of IRF6, CBL and IFNB1 in AML-193 cells infected with AL645608.3 overexpression lentiviruses. ns, P>0.05; ****P<0.0001.

Article Snippet: The transferred membranes were blocked for 1 h. After discarding the sealing liquid, the diluted primary antibodies against IRF6 (#sc-37704; 1:500; Santa, Texas, USA), CBL (#25818-1-AP; 1:1000; Proteintech, Wuhan, China), IFNB1 (#ab275880; 1:500; Abcam, Massachusetts, USA), and GAPDH (#ab181602; 1:10000; Abcam) were added and incubated overnight at 4°C.

Techniques: Expressing, Infection, Over Expression

Journal: American Journal of Translational Research

Article Title: LncRNA AL645608.3 mediates malignant progression of acute myeloid leukemia

doi:

Figure Lengend Snippet: Effects of AL645608.3 on the protein expression of IRF6, CBL and IFNB1 in AML cells. A. Representative western blot images for IRF6, CBL and IFNB1 in THP-1 cells with sh-AL645608.3 lentiviruses infection and in AML-193 cells with AL645608.3 overexpression lentiviruses infection. B-D. Western blot for the detection of protein levels of IRF6, CBL and IFNB1 in THP-1 cells infected with sh-AL645608.3 lentiviruses. E-G. Western blot for the detection of protein levels of IRF6, CBL and IFNB1 in AML-193 cells infected with AL645608.3 overexpression lentiviruses. ns, P>0.05; **P<0.01; ***P<0.001; ****P<0.0001.

Article Snippet: The transferred membranes were blocked for 1 h. After discarding the sealing liquid, the diluted primary antibodies against IRF6 (#sc-37704; 1:500; Santa, Texas, USA), CBL (#25818-1-AP; 1:1000; Proteintech, Wuhan, China), IFNB1 (#ab275880; 1:500; Abcam, Massachusetts, USA), and GAPDH (#ab181602; 1:10000; Abcam) were added and incubated overnight at 4°C.

Techniques: Expressing, Western Blot, Infection, Over Expression

Journal: Journal of Neuroinflammation

Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice

doi: 10.1186/s12974-017-1039-7

Figure Lengend Snippet: Expressions of DRD1 and IFN-beta receptor in microglia. a Co-localization of DRD1 with Iba 1 and of IFN-beta receptor with Iba 1 was observed in the perihematoma brain tissue (showing with black triangle) at 24 h after ICH. b Representative photographs of co-localization of DRD1 with Iba 1 (bar = 50 μm). c Representative photographs of co-localization of IFN-beta receptor with Iba 1 (bar = 50 μm)

Article Snippet: After blocking with 5% nonfat milk for 1.5 h, the membranes were incubated overnight at 4 °C with the primary antibodies: rabbit anti-DRD1 (1:1000, Abcam), goat anti-IFN-beta (1:300, Santa Cruz), rabbit anti-IFN-beta receptor (1:500, Santa Cruz), rabbit anti-STAT1(1:2000, Cell Signaling), goat anti-p-STAT1 (1:500, Santa Cruz), rabbit anti-NLRP3 (1:1000, MyBioSource), rabbit anti-caspase1 (1:1000, NOVUS Biologicals), rabbit anti-IL-1beta (1:2000, Cell Signaling), and rabbit anti-β-actin (1:2000, Santa Cruz). β-actin served as the loading control.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice

doi: 10.1186/s12974-017-1039-7

Figure Lengend Snippet: Expressions of DRD1 and IFN-beta in the ipsilateral hemispheres after intracerebral hemorrhage (ICH). a Representative photographs of DRD1 and IFN-beta in western blotting. b Bar graphs of quantitative analysis of DRD1 and IFN-beta expressions from the ipsilateral hemisphere after ICH. DRD1 and IFN-beta expressions increased, * p < 0.05, ** p < 0.01 versus sham; DRD1 and IFN-beta expressions decreased, # p < 0.05, ## p < 0.01 versus sham

Article Snippet: After blocking with 5% nonfat milk for 1.5 h, the membranes were incubated overnight at 4 °C with the primary antibodies: rabbit anti-DRD1 (1:1000, Abcam), goat anti-IFN-beta (1:300, Santa Cruz), rabbit anti-IFN-beta receptor (1:500, Santa Cruz), rabbit anti-STAT1(1:2000, Cell Signaling), goat anti-p-STAT1 (1:500, Santa Cruz), rabbit anti-NLRP3 (1:1000, MyBioSource), rabbit anti-caspase1 (1:1000, NOVUS Biologicals), rabbit anti-IL-1beta (1:2000, Cell Signaling), and rabbit anti-β-actin (1:2000, Santa Cruz). β-actin served as the loading control.

Techniques: Western Blot

Journal: Journal of Neuroinflammation

Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice

doi: 10.1186/s12974-017-1039-7

Figure Lengend Snippet: Effects of A68930 on the expressions of proteins in DRD1/IFN-beta/STAT1/NLRP3 pathway after ICH. a Representative photographs of the expressions of proteins in DRD1/IFN-beta/STAT1/NLRP3 pathway in western blotting. b and c Bar graphs of quantitative analysis of DRD1, IFN-beta, IFN-beta receptor, p-STAT1, NLRP3, pro-caspase1, caspase1, pro-IL-1beta, and IL-1beta, respectively. * p < 0.05, ** p < 0.01 versus sham; # p < 0.05, ## p < 0.01 versus vehicle

Article Snippet: After blocking with 5% nonfat milk for 1.5 h, the membranes were incubated overnight at 4 °C with the primary antibodies: rabbit anti-DRD1 (1:1000, Abcam), goat anti-IFN-beta (1:300, Santa Cruz), rabbit anti-IFN-beta receptor (1:500, Santa Cruz), rabbit anti-STAT1(1:2000, Cell Signaling), goat anti-p-STAT1 (1:500, Santa Cruz), rabbit anti-NLRP3 (1:1000, MyBioSource), rabbit anti-caspase1 (1:1000, NOVUS Biologicals), rabbit anti-IL-1beta (1:2000, Cell Signaling), and rabbit anti-β-actin (1:2000, Santa Cruz). β-actin served as the loading control.

Techniques: Western Blot

Journal: Journal of Neuroinflammation

Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice

doi: 10.1186/s12974-017-1039-7

Figure Lengend Snippet: DRD1 antagonist and IFN-beta siRNA reversed effects of A68930 on neurological outcome and brain water content after ICH. Garcia test score ( a ), left forelimb placement ( b ), and brain water content ( c ) at 24 h after ICH. ** p < 0.01 versus sham; # p < 0.05, ## p < 0.01 versus vehicle; $ p < 0.05, $$ p < 0.01 versus A68930 group; & p < 0.05 versus scramble siRNA group

Article Snippet: After blocking with 5% nonfat milk for 1.5 h, the membranes were incubated overnight at 4 °C with the primary antibodies: rabbit anti-DRD1 (1:1000, Abcam), goat anti-IFN-beta (1:300, Santa Cruz), rabbit anti-IFN-beta receptor (1:500, Santa Cruz), rabbit anti-STAT1(1:2000, Cell Signaling), goat anti-p-STAT1 (1:500, Santa Cruz), rabbit anti-NLRP3 (1:1000, MyBioSource), rabbit anti-caspase1 (1:1000, NOVUS Biologicals), rabbit anti-IL-1beta (1:2000, Cell Signaling), and rabbit anti-β-actin (1:2000, Santa Cruz). β-actin served as the loading control.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice

doi: 10.1186/s12974-017-1039-7

Figure Lengend Snippet: DRD1 antagonist and IFN-beta siRNA reversed effects of A68930 on DRD1/IFN-beta/STAT1/NLRP3 signaling pathway after ICH. a Representative photographs of the expressions of proteins in DRD1/IFN-beta/STAT1/NLRP3 pathway in western blotting. b and c Bar graphs of quantitative analysis of DRD1, IFN-beta, IFN-beta receptor, p-STAT1, NLRP3, pro-caspase1, caspase1, pro-IL-1beta, and IL-1beta, respectively. * p < 0.05, ** p < 0.01 versus sham; # p < 0.05, ## p < 0.01 versus vehicle; $ p < 0.05, $$ p < 0.01 versus A68930 group; & p < 0.05, && p < 0.01 versus scramble siRNA group

Article Snippet: After blocking with 5% nonfat milk for 1.5 h, the membranes were incubated overnight at 4 °C with the primary antibodies: rabbit anti-DRD1 (1:1000, Abcam), goat anti-IFN-beta (1:300, Santa Cruz), rabbit anti-IFN-beta receptor (1:500, Santa Cruz), rabbit anti-STAT1(1:2000, Cell Signaling), goat anti-p-STAT1 (1:500, Santa Cruz), rabbit anti-NLRP3 (1:1000, MyBioSource), rabbit anti-caspase1 (1:1000, NOVUS Biologicals), rabbit anti-IL-1beta (1:2000, Cell Signaling), and rabbit anti-β-actin (1:2000, Santa Cruz). β-actin served as the loading control.

Techniques: Western Blot

Journal: eLife

Article Title: LRRK2 maintains mitochondrial homeostasis and regulates innate immune responses to Mycobacterium tuberculosis

doi: 10.7554/eLife.51071

Figure Lengend Snippet:

Article Snippet: Antibody , anti-IFNB Rabbit polyclonal , PBL Assay Science , 32400–1 , (1:250 neutralizing).

Techniques: Sequencing, Recombinant

Journal: Journal of Neuroinflammation

Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice

doi: 10.1186/s12974-017-1039-7

Figure Lengend Snippet: Expressions of DRD1 and IFN-beta receptor in microglia. a Co-localization of DRD1 with Iba 1 and of IFN-beta receptor with Iba 1 was observed in the perihematoma brain tissue (showing with black triangle) at 24 h after ICH. b Representative photographs of co-localization of DRD1 with Iba 1 (bar = 50 μm). c Representative photographs of co-localization of IFN-beta receptor with Iba 1 (bar = 50 μm)

Article Snippet: A series of coronal brain sections (10 μm thick) were blocked in 5% bovine serum albumin for 2 h at room temperature and then were incubated with rabbit anti-DRD1 (1:200, Abcam), rabbit anti-IFN-beta receptor (1:50, Santa Cruz), goat anti-ionized calcium-binding adapter molecule 1 (Iba 1, 1:200, Abcam), and rabbit anti-MPO (1:100, Abcam) at 4 °C overnight.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice

doi: 10.1186/s12974-017-1039-7

Figure Lengend Snippet: Expressions of DRD1 and IFN-beta in the ipsilateral hemispheres after intracerebral hemorrhage (ICH). a Representative photographs of DRD1 and IFN-beta in western blotting. b Bar graphs of quantitative analysis of DRD1 and IFN-beta expressions from the ipsilateral hemisphere after ICH. DRD1 and IFN-beta expressions increased, * p < 0.05, ** p < 0.01 versus sham; DRD1 and IFN-beta expressions decreased, # p < 0.05, ## p < 0.01 versus sham

Article Snippet: A series of coronal brain sections (10 μm thick) were blocked in 5% bovine serum albumin for 2 h at room temperature and then were incubated with rabbit anti-DRD1 (1:200, Abcam), rabbit anti-IFN-beta receptor (1:50, Santa Cruz), goat anti-ionized calcium-binding adapter molecule 1 (Iba 1, 1:200, Abcam), and rabbit anti-MPO (1:100, Abcam) at 4 °C overnight.

Techniques: Western Blot

Journal: Journal of Neuroinflammation

Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice

doi: 10.1186/s12974-017-1039-7

Figure Lengend Snippet: Effects of A68930 on the expressions of proteins in DRD1/IFN-beta/STAT1/NLRP3 pathway after ICH. a Representative photographs of the expressions of proteins in DRD1/IFN-beta/STAT1/NLRP3 pathway in western blotting. b and c Bar graphs of quantitative analysis of DRD1, IFN-beta, IFN-beta receptor, p-STAT1, NLRP3, pro-caspase1, caspase1, pro-IL-1beta, and IL-1beta, respectively. * p < 0.05, ** p < 0.01 versus sham; # p < 0.05, ## p < 0.01 versus vehicle

Article Snippet: A series of coronal brain sections (10 μm thick) were blocked in 5% bovine serum albumin for 2 h at room temperature and then were incubated with rabbit anti-DRD1 (1:200, Abcam), rabbit anti-IFN-beta receptor (1:50, Santa Cruz), goat anti-ionized calcium-binding adapter molecule 1 (Iba 1, 1:200, Abcam), and rabbit anti-MPO (1:100, Abcam) at 4 °C overnight.

Techniques: Western Blot

Journal: Journal of Neuroinflammation

Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice

doi: 10.1186/s12974-017-1039-7

Figure Lengend Snippet: DRD1 antagonist and IFN-beta siRNA reversed effects of A68930 on neurological outcome and brain water content after ICH. Garcia test score ( a ), left forelimb placement ( b ), and brain water content ( c ) at 24 h after ICH. ** p < 0.01 versus sham; # p < 0.05, ## p < 0.01 versus vehicle; $ p < 0.05, $$ p < 0.01 versus A68930 group; & p < 0.05 versus scramble siRNA group

Article Snippet: A series of coronal brain sections (10 μm thick) were blocked in 5% bovine serum albumin for 2 h at room temperature and then were incubated with rabbit anti-DRD1 (1:200, Abcam), rabbit anti-IFN-beta receptor (1:50, Santa Cruz), goat anti-ionized calcium-binding adapter molecule 1 (Iba 1, 1:200, Abcam), and rabbit anti-MPO (1:100, Abcam) at 4 °C overnight.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice

doi: 10.1186/s12974-017-1039-7

Figure Lengend Snippet: DRD1 antagonist and IFN-beta siRNA reversed effects of A68930 on DRD1/IFN-beta/STAT1/NLRP3 signaling pathway after ICH. a Representative photographs of the expressions of proteins in DRD1/IFN-beta/STAT1/NLRP3 pathway in western blotting. b and c Bar graphs of quantitative analysis of DRD1, IFN-beta, IFN-beta receptor, p-STAT1, NLRP3, pro-caspase1, caspase1, pro-IL-1beta, and IL-1beta, respectively. * p < 0.05, ** p < 0.01 versus sham; # p < 0.05, ## p < 0.01 versus vehicle; $ p < 0.05, $$ p < 0.01 versus A68930 group; & p < 0.05, && p < 0.01 versus scramble siRNA group

Article Snippet: A series of coronal brain sections (10 μm thick) were blocked in 5% bovine serum albumin for 2 h at room temperature and then were incubated with rabbit anti-DRD1 (1:200, Abcam), rabbit anti-IFN-beta receptor (1:50, Santa Cruz), goat anti-ionized calcium-binding adapter molecule 1 (Iba 1, 1:200, Abcam), and rabbit anti-MPO (1:100, Abcam) at 4 °C overnight.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Nucleosomes Are Stably Evicted from Enhancers but Not Promoters upon Induction of Certain Pro-Inflammatory Genes in Mouse Macrophages

doi: 10.1371/journal.pone.0093971

Figure Lengend Snippet: (A and B), Nucleosome occupancy at an enhancer 10 kb upstream of the TSS of IL12B in BMDMs was analyzed before induction (blue bars and lines), and after 1.5 h (yellow), 3 h (orange), 5 h (light red) and 10 h (dark red) of growth of cells in the presence of 1 μg/ml LPS, using the assay described in with modifications detailed in the . In brief, occupancy was measured by determining the sensitivity of cross-linked chromatin to a wide range of MNase. Digestion data for each genomic location analyzed by qRT-PCR with specific primer pairs was fitted to two-state exponential decay functions and the percentage of DNA in the population of cells found to be protected against MNase by binding of a nucleosome is indicated on the y-axis. In panel (A) each overlapping colored bar represents the length of the amplicon measured. The minimal enhancer that was shown by Zhou et al. to contain the LPS-inducible DNaseI hypersensitive site HSS1 as well as consensus-sites for Oct1/2 and C/EBPβ is indicated by the black bar . Consensus-sites for PU.1, NFκB, AP1 and IRF identified using ConSite are indicated. In panel (B) nucleosome occupancy at the midpoint of each amplicon measured by the experiment performed in panel (A) is indicated by a dot, with error bars showing the SEM of at least two independent measurements (10 h was measured only once). (C and D), BMDMs were induced as described in (A) and nucleosome occupancy in a region surrounding the TSS of IL12B was determined. The data is displayed as in panels (A) and (B) respectively. The black bar below the data in (C) indicates the TSS and the light blue bars indicate putative TATA-boxes predicted by ConSite. (E), Expression of IL12B, IFNB1 and IL1A in response to LPS. mRNA from BMDMs induced with LPS as in panel A as well as from splenic B-cells was isolated as described in the , reverse transcribed and cDNA quantified by qRT-PCR. Data was normalized to a location in the ORF of the constitutively expressed RPL4 gene. The SEM of at least two independent measurements is indicated (10 h timepoint was measured only once).

Article Snippet: In contrast to our findings at the IL1A and IL12B promoters we found that the TATAA-sequence in the IFNB1 promoter was cleared of nucleosomes upon induction in primary mouse macrophages as had been described for the IFNB1 promoter in human cells ( ) .

Techniques: Quantitative RT-PCR, Binding Assay, Amplification, Expressing, Isolation, Reverse Transcription

Journal: PLoS ONE

Article Title: Nucleosomes Are Stably Evicted from Enhancers but Not Promoters upon Induction of Certain Pro-Inflammatory Genes in Mouse Macrophages

doi: 10.1371/journal.pone.0093971

Figure Lengend Snippet: (A and B), Nucleosome occupancy was determined in BMDMs before induction (blue bars and lines), and upon induction with 1 μg/ml LPS for 1.5 h with (green) or without (yellow) pretreatment of cells with 1 μM TPG for 1 h. The minimal enhancer region (black bar) and binding sites for XBP, AP1, IRF3 and NFκB identified by are shown in (A). ConSite predicted binding sites for PU.1 and C/EBP are indicated. (C and D), Nucleosome occupancy at the proximal enhancer and promoter of IFNB1 was determined as in panel A and analyzed in a region encompassing the proximal enhancer that is conserved in humans and has been shown to form an enhanceosome upon viral stimulation of HeLa cells , as well as in the 5′ region of the IFNB1 ORF. Conserved binding sites for NFκB, ATF, AP1 and IRF3/7 identified by are indicated. ConSite-predicted consensus sites for PU.1 and C/EBP are also shown. E , Expression of IFNB1 upon stimulation with different inducers. BMDMs were induced for 3 h (dark blue) or 16 h (light blue) with 1 μg/ml of LPS, or 1 μg/ml of ISD, 1 μg/ml p(I:C), or 1 μg/ml p(dA:dT) added either alone or together with LPS as indicated. Where indicated cells were pre-treated with the ER-stress inducer TPG for 1 h prior to LPS induction. mRNA was isolated and quantified as described in the . Data was normalized to the ORF of RPL4.

Article Snippet: In contrast to our findings at the IL1A and IL12B promoters we found that the TATAA-sequence in the IFNB1 promoter was cleared of nucleosomes upon induction in primary mouse macrophages as had been described for the IFNB1 promoter in human cells ( ) .

Techniques: Binding Assay, Expressing, Isolation