Journal: The Journal of Physiology

Article Title: Sprint interval and traditional endurance training increase net intramuscular triglyceride breakdown and expression of perilipin 2 and 5

doi: 10.1113/jphysiol.2012.240952

Figure Lengend Snippet: Representative immunofluorescence images of PLIN5 (A), IMTG (B), and the merged images (C) with the yellow areas describing regions of overlap between PLIN5 and IMTG images. The yellow objects were then extracted (to measure the number of PLIN5-LDs; D), and the number subtracted from the total number of LDs to quantify the number of PLIN5-null-LD. The same procedure was used to obtain values of colocalization between PLIN2 and IMTG. White bar, 5 μm. The number of PLIN2-LD (filled bars) and PLIN2-null-LD (open bars) were quantified before (PreEx) and after exercise (PostEx), prior to (E) and following (F) training in both the SIT and ET groups. The same analysis was repeated for PLIN5 for pre- (G) and post-training (H). *Main effect of exercise bout (P < 0.05 vs. pre-exercise).

Article Snippet: The specificity of the PLIN5 antibody was confirmed in a competition experiment in which a PLIN5 recombinant peptide (Cat. no. NB110–60511PEP; Novus Biologicals, Cambridge, UK) was incubated with the PLIN5 primary antibody, subsequently resulting in removal of the positive fluorescent signal for PLIN5 (Supplementary Fig. S1 ).

Techniques: Immunofluorescence

Journal: The Journal of Physiology

Article Title: Sprint interval and traditional endurance training increase net intramuscular triglyceride breakdown and expression of perilipin 2 and 5

doi: 10.1113/jphysiol.2012.240952

Figure Lengend Snippet: Representative immunofluorescence images of PLIN5 (stained green) in combination with WGA to identify the cell border (stained blue) in skeletal muscle, pre- (A) and post- (B) 6 weeks of SIT. Type I fibres are indicated with a ‘I’, all other fibres are assumed type II fibres. White bar, 50 μm. PLIN5 expression, quantified from immunofluorescence images of PLIN5, in type I and type II fibres obtained before (C) and after (D) 6 weeks of SIT (filled bars) or ET (open bars). PLIN5 expression quantified from immunofluorescence images correlated with PLIN5 expression determined following immunoblotting of whole muscle homogenates (E). Values are presented as means ± SEM (n= 8 per group). *Main effect of fibre type (P < 0.05 vs. type I fibres). †Main effect of training intervention (P < 0.05 vs. pre-training).

Article Snippet: The specificity of the PLIN5 antibody was confirmed in a competition experiment in which a PLIN5 recombinant peptide (Cat. no. NB110–60511PEP; Novus Biologicals, Cambridge, UK) was incubated with the PLIN5 primary antibody, subsequently resulting in removal of the positive fluorescent signal for PLIN5 (Supplementary Fig. S1 ).

Techniques: Immunofluorescence, Staining, Expressing, Western Blot

Journal: The Journal of Physiology

Article Title: Sprint interval and traditional endurance training increase net intramuscular triglyceride breakdown and expression of perilipin 2 and 5

doi: 10.1113/jphysiol.2012.240952

Figure Lengend Snippet: Bivariate correlation analysis

Article Snippet: The specificity of the PLIN5 antibody was confirmed in a competition experiment in which a PLIN5 recombinant peptide (Cat. no. NB110–60511PEP; Novus Biologicals, Cambridge, UK) was incubated with the PLIN5 primary antibody, subsequently resulting in removal of the positive fluorescent signal for PLIN5 (Supplementary Fig. S1 ).

Techniques:

Journal: PLoS ONE

Article Title: Immunohistochemical, Ultrastructural and Functional Analysis of Axonal Regeneration through Peripheral Nerve Grafts Containing Schwann Cells Expressing BDNF, CNTF or NT3

doi: 10.1371/journal.pone.0069987

Figure Lengend Snippet: (D) Quantification revealed significantly greater numbers of IB 4 + axons in normal nerves compared to acellular grafts (*) and in NT3 grafts compared to all other experimental groups (**). (E) The number of CGRP + axons was not significantly different between experimental groups. Values represent M ± SEM of n = 3; p<0.05. Further details on statistical analysis provided as . Scale bar for A–C: 100 µm.

Article Snippet: Immunostaining was done at room temperature for antibodies to calcitonin gene-related peptide (CGRP) (AbDSerotec #1720-9007, 1∶1000) to label axons of primary peptidergic sensory neurons.

Techniques:

Journal: PLoS ONE

Article Title: Immunohistochemical, Ultrastructural and Functional Analysis of Axonal Regeneration through Peripheral Nerve Grafts Containing Schwann Cells Expressing BDNF, CNTF or NT3

doi: 10.1371/journal.pone.0069987

Figure Lengend Snippet: Summary of data.

Article Snippet: Immunostaining was done at room temperature for antibodies to calcitonin gene-related peptide (CGRP) (AbDSerotec #1720-9007, 1∶1000) to label axons of primary peptidergic sensory neurons.

Techniques: Functional Assay

Journal: Scientific Reports

Article Title: S tellera chamaejasme and its constituents induce cutaneous wound healing and anti-inflammatory activities

doi: 10.1038/srep42490

Figure Lengend Snippet: Human dermal fibroblasts (Hs68) were incubated for 48 h with SCE (1,5, or 10 μg/ml) or various compounds (10 or 20 μM). ( A , B ) mRNA levels of COL1A1, COL3A1 were determined with RT-PCR. ( C , D ) Production of type I procollagen was determined with ELISA. Results are expressed as the mean ± SD of three independent experiments. ( ## p < 0.01 and ** p < 0.01).

Article Snippet: Cell culture medium was collected after 24 h, type I procollagen production were quantified using a procollagen type I C-peptide enzyme immunoassay kit (MK101; Takara, Shiga, Japan).

Techniques: Incubation, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: The cytoplasmic nuclear receptor RARγ controls RIP1 initiated cell death when cIAP activity is inhibited

doi: 10.1038/s41467-017-00496-6

Figure Lengend Snippet: RARγ has a role in the formation of death complexes. a Western blot analysis of HT-29 cont-shRNA, and RARγ-shRNA-A treated with DMSO, TS or TSZ for 24 h and cell lysates were immunoblotted with the indicated antibodies. b , c Immunoprecipitation of HT-29 cont-shRNA or RARγ-shRNA-A cells treated with TSZ for the indicated time. Cell lysates were collected and immunoprecipitated with anti-Caspase-8 antibody b or anti-TNFR1 antibody c . The immunoprecipitated complexes were immunoblotted with the indicated antibodies. d Sequential immunoprecipitation of HT-29 cont-shRNA or RARγ-shRNA-A cells treated TSZ for 2 h. First IP : TNFR1 complex I was immunoprecipitated using anti-TNFR1 antibody. Second IP : the remaining lysates were immunoprecipitated again with anti-TNFR1 antibody. Third IP : the remaining lysates were then immunoprecipitated with anti-TRADD antibody. The immunoprecipitated complexes were analyzed by with the indicated antibodies. (M: marker). All blots above are representative of one of three experiments

Article Snippet: Anti-RARγ (C-15) (sc-550) for human, anti-RARα (C-20) (sc-551), anti-caspase-8 (C-20) (sc-6136), anti-cIAP2 (sc7944) and anti-Fas (C-20) (sc-715) from Santa Cruz; anti-RIP1 (610459) and anti-FADD (610400) from BD Biosciences; anti-RARγ1 (ab5905) for mouse, anti-RIP3 (ab72106), anti-MLKL (ab184718) for human, anti-MLKL (ab172868) for mouse and anti-cIAP1 (ab2399) from Abcam; anti-RIP3 (2283) for mouse from ProSci, anti-TRADD (05-473) from Upstate; anti-TRAF2 (MAB3277) and anti-TNFR1 (AF-425-PB) from R&D; anti-Actin (A3853) (dilution 1:10,000), anti-FLAG (F9291) (dilution 1:5,000) and anti-GFP (G6539) (dilution 1:5 000) from Sigma; anti-V5 (R960-25) (dilution 1:5,000) from Invitrogen; anti-PARP1 (BML-SA250-0050) from Enzo Life Science; anti-GAPDH (NB300-22) (dilution 1:5,000) from Novus Biologicals; anti-cleaved caspase-8 (9496), anti-CYLD (4495), anti-RIP1 (137451) and anti-p-RIP1(65746) from Cell Signaling Technology; anti-DsRed (632392) (dilution 1:5,000) from Clontech.

Techniques: Western Blot, shRNA, Immunoprecipitation, Marker

Journal: Nature Communications

Article Title: The cytoplasmic nuclear receptor RARγ controls RIP1 initiated cell death when cIAP activity is inhibited

doi: 10.1038/s41467-017-00496-6

Figure Lengend Snippet: RARγ initiates the formation of death complexes by dissociating RIP1 from TNFR1. a Immunoprecipitation of HT-29 cont-shRNA, TRADD-shRNA or RIP1-shRNA treated with TSZ for the indicated times. Cell lysates were immunoprecipitated using anti-TNFR1 antibody and immunoblotted with the indicated antibodies. b Sequential immunoprecipitation of HEK293T cells co-transfected with FLAG-TNFR1, DsRed-RIP1 and increasing amounts of V5-RARγ-NLSmut plasmids as indicated. First IP : cell lysates were immunoprecipitated with anti-FLAG (TNFR1) antibody. Second IP : the remaining lysates were then immunoprecipitated with anti-V5 (RARγ) antibody. The immunoprecipitated complexes were analyzed by with the indicated antibodies. c Sequential immunoprecipitation of HEK293T cells co-transfected with V5-RARγ-NLSmut, RIP1-Myc, and increasing amounts of DsRed-RIP3 plasmids as indicated. First IP : cell lysates were immunoprecipitated with anti-V5 (RARγ) antibody. Second IP : the remaining lysates were then immunoprecipitated with anti-DsRed (RIP3) antibody. The immunoprecipitated complexes were analyzed with the indicated antibodies. d WT and RIP3−/− MEFs treated with TSZ for the indicated times. Cell lysates were immunoprecipitated using anti-Caspase-8 antibody and immunoblotted with the indicated antibodies. All blots above are representative of one of three experiments

Article Snippet: Anti-RARγ (C-15) (sc-550) for human, anti-RARα (C-20) (sc-551), anti-caspase-8 (C-20) (sc-6136), anti-cIAP2 (sc7944) and anti-Fas (C-20) (sc-715) from Santa Cruz; anti-RIP1 (610459) and anti-FADD (610400) from BD Biosciences; anti-RARγ1 (ab5905) for mouse, anti-RIP3 (ab72106), anti-MLKL (ab184718) for human, anti-MLKL (ab172868) for mouse and anti-cIAP1 (ab2399) from Abcam; anti-RIP3 (2283) for mouse from ProSci, anti-TRADD (05-473) from Upstate; anti-TRAF2 (MAB3277) and anti-TNFR1 (AF-425-PB) from R&D; anti-Actin (A3853) (dilution 1:10,000), anti-FLAG (F9291) (dilution 1:5,000) and anti-GFP (G6539) (dilution 1:5 000) from Sigma; anti-V5 (R960-25) (dilution 1:5,000) from Invitrogen; anti-PARP1 (BML-SA250-0050) from Enzo Life Science; anti-GAPDH (NB300-22) (dilution 1:5,000) from Novus Biologicals; anti-cleaved caspase-8 (9496), anti-CYLD (4495), anti-RIP1 (137451) and anti-p-RIP1(65746) from Cell Signaling Technology; anti-DsRed (632392) (dilution 1:5,000) from Clontech.

Techniques: Immunoprecipitation, shRNA, Transfection

Journal: Nature Communications

Article Title: The cytoplasmic nuclear receptor RARγ controls RIP1 initiated cell death when cIAP activity is inhibited

doi: 10.1038/s41467-017-00496-6

Figure Lengend Snippet: RARγ modulates TNF-induced RIP-initiated cell death. a TRADD is responsible for initiated TC or TCZ-induced cell death. b RARγ is essential for the transition from inflammatory signaling to death pathways in TNF-induced RIP1-initiated cell death. When RARγ is released from the nucleus in response to TNF under the conditions of cIAP inhibition, it initiates the formation of death complex IIa by dissociating RIP1 from TNFR1. When RIP1 recruits RIP3 to necrosome/complex IIb, RARγ is replaced from the complex

Article Snippet: Anti-RARγ (C-15) (sc-550) for human, anti-RARα (C-20) (sc-551), anti-caspase-8 (C-20) (sc-6136), anti-cIAP2 (sc7944) and anti-Fas (C-20) (sc-715) from Santa Cruz; anti-RIP1 (610459) and anti-FADD (610400) from BD Biosciences; anti-RARγ1 (ab5905) for mouse, anti-RIP3 (ab72106), anti-MLKL (ab184718) for human, anti-MLKL (ab172868) for mouse and anti-cIAP1 (ab2399) from Abcam; anti-RIP3 (2283) for mouse from ProSci, anti-TRADD (05-473) from Upstate; anti-TRAF2 (MAB3277) and anti-TNFR1 (AF-425-PB) from R&D; anti-Actin (A3853) (dilution 1:10,000), anti-FLAG (F9291) (dilution 1:5,000) and anti-GFP (G6539) (dilution 1:5 000) from Sigma; anti-V5 (R960-25) (dilution 1:5,000) from Invitrogen; anti-PARP1 (BML-SA250-0050) from Enzo Life Science; anti-GAPDH (NB300-22) (dilution 1:5,000) from Novus Biologicals; anti-cleaved caspase-8 (9496), anti-CYLD (4495), anti-RIP1 (137451) and anti-p-RIP1(65746) from Cell Signaling Technology; anti-DsRed (632392) (dilution 1:5,000) from Clontech.

Techniques: Inhibition