Journal: PLoS ONE

Article Title: Oil degradation potential of microbial communities in water and sediment of Baltic Sea coastal area

doi: 10.1371/journal.pone.0218834

Figure Lengend Snippet: Copy numbers determined by qPCR from triplicate DNA extractions from (A) sea water (mL -1 ) and (B) untreated sediment or sediment treated with DNase (g -1 dry weight).

Article Snippet: The reactions consisted of 13.8 μl RNA extract, 1.7 μl 10x reaction buffer and 1.7 μl DNase RQ1 (1000 U mL -1 , Promega, WI, USA) and were incubated at 37°C for 30 min, after which the DNase reaction was inactivated by adding 1.8 μl DNase Stop Solution followed by a final 10 min incubation at 65°C.

Techniques:

Journal: PLoS ONE

Article Title: Oil degradation potential of microbial communities in water and sediment of Baltic Sea coastal area

doi: 10.1371/journal.pone.0218834

Figure Lengend Snippet: Copy numbers determined by qPCR from triplicate DNA and RNA sample extractions isolated from A) sea water (mL -1 ) B) untreated sediment or sediment treated with DNase (g -1 dry weight).

Article Snippet: The reactions consisted of 13.8 μl RNA extract, 1.7 μl 10x reaction buffer and 1.7 μl DNase RQ1 (1000 U mL -1 , Promega, WI, USA) and were incubated at 37°C for 30 min, after which the DNase reaction was inactivated by adding 1.8 μl DNase Stop Solution followed by a final 10 min incubation at 65°C.

Techniques: Isolation

Journal: PLoS ONE

Article Title: Oil degradation potential of microbial communities in water and sediment of Baltic Sea coastal area

doi: 10.1371/journal.pone.0218834

Figure Lengend Snippet: Bacteria detected from A) RNA, B) DNA isolated from the sea water samples, C) DNA isolated from the DNase treated sediment samples and D) DNA extracted directly from the sediment samples.

Article Snippet: The reactions consisted of 13.8 μl RNA extract, 1.7 μl 10x reaction buffer and 1.7 μl DNase RQ1 (1000 U mL -1 , Promega, WI, USA) and were incubated at 37°C for 30 min, after which the DNase reaction was inactivated by adding 1.8 μl DNase Stop Solution followed by a final 10 min incubation at 65°C.

Techniques: Bacteria, Isolation

Journal: PLoS ONE

Article Title: Oil degradation potential of microbial communities in water and sediment of Baltic Sea coastal area

doi: 10.1371/journal.pone.0218834

Figure Lengend Snippet: Archaea detected from A) RNA, B) DNA isolated from the sea water samples and C) DNA isolated from the DNase treated sediment samples and D) the DNA extracted directly from the sediment samples.

Article Snippet: The reactions consisted of 13.8 μl RNA extract, 1.7 μl 10x reaction buffer and 1.7 μl DNase RQ1 (1000 U mL -1 , Promega, WI, USA) and were incubated at 37°C for 30 min, after which the DNase reaction was inactivated by adding 1.8 μl DNase Stop Solution followed by a final 10 min incubation at 65°C.

Techniques: Isolation

Journal: Scientific Reports

Article Title: Mitochondrial Ca 2+ uptake controls actin cytoskeleton dynamics during cell migration

doi: 10.1038/srep36570

Figure Lengend Snippet: ( a ) Representative confocal images showing the accumulation of phospho-myosin light chain (P-MLC) signal in F-actin fibers in sh2 MCU cells compared to controls. P-MLC and F-actin were stained using P-MLC antibody and phalloidine rhodamine probe, respectively. Merge channels between P-MLC (green) and F-actin (red) were presented in the rightmost panels. Scale bar: 20 μm. ( b ) Immunoblot showing the increase of MLC phosphorylation on Serine19 (P-MLC) and Threonine18 and Serine19 (P 2 -MLC) positions in mcu -silenced cells. MCU antibody was used to show the efficiency of the shRNAs. F 1 F 0 ATPase antibody was used as loading control. ( c ) Representative confocal images showing the accumulation of Vinculin in FAPs in sh2 MCU cells compared to control. Vinculin and F-actin were detected using anti-Vinculin antibody and phalloidine rhodamine probe, respectively. Merge channels between Vinculin (green) and F-actin (red) were presented in the rightmost panels. Scale bar: 20 μm. ( d ) Representative time-lapse confocal images showing the persistence of Paxillin-GFP signal in the FAPs of mcu -knockdown cells compared to control. Scale bar: 5 μm. ( e ) Histogram representing the increase of FAPs number estimated from Vinculin staining in sh2 MCU cells compared to control. (mean ± S.E.M; 10 cells for each condition; n = 3 independent experiments). ***P < 0.001. ( f ) Histogram representing the increase of FAP size estimated from Vinculin staining in sh2 MCU cells compared to control cells. (mean ± S.E.M; 10 cells for each condition; n = 3 independent experiments). ***P < 0.001. ( g ) Immunoblot showing the effect of sh1 and sh2 MCU on phospho-Paxillin Y118 protein levels (P-Paxillin Y118). Quantitative ratios between P-Paxillin Y118 and total Paxillin are indicated below. Mcu knockdown leads to decreased P-Paxillin Y118/total Paxillin ratio. MCU antibody was used to show the efficiency of the shRNAs. F 1 F 0 ATPase antibody was used as loading control. ( h ) Histogram presenting the decrease of Calpain activity measured in mcu -silenced cells and compared to control cells (mean ± S.E.M; n = 3 independent experiments). *P < 0.05, **P < 0.01. See also .

Article Snippet: The following antibodies were used: MCU (Abcam #Ab121499; dilution: 1/500), F 1 F 0 ATPase (BD #612518; dilution: 1/1000), Vinculin (Santa Cruz #sc-55465; dilution: 1/2000), Paxillin H114 (Santa Cruz #sc-5574; dilution: 1/1000), Phospho-Paxillin Tyr 118 (Cell Signaling Technology #2541; dilution: 1/1000), Phospho-Myosin light chain 2 (Cell Signaling Technology #3671; dilution: 1/1000), STIM1 (Abcam #Ab57834; dilution: 1/1000) and Rac1 (Cell Biolabs #240106; dilution: 1/1000).

Techniques: Staining, Western Blot, Activity Assay