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It’s a visual representation of just how many balloons there are floating through the sky:
Image: The Nowhere Twins – YouTube
Now that’s a lot of balloons. These balloons are probably adorably scrunched and hidden underneath a blanket on a bed in North Carolina or Missouri (at least we hope so), where the Nowhere Twins are finding them.
Image: The Nowhere Twins – YouTube
In the video, twin girls, who are named as Lilly and Sophie, lie on a blanket in a swamp. They’re so excited to find this windblown balloon stash that they jump up and down like crazy.
Image: The Nowhere Twins – YouTube
Because their mom is video-recording them, you get to see the twins’ reaction to discovering a bunch of pink and white balloons.
Image: The Nowhere Twins – YouTube
This has got to be one of the cutest “sibling” moments ever on YouTube, and it’s already been viewed over one million times. It’s hard to tell, but they’re saying “Daddy!!” in the background, as well.
Image: The Nowhere Twins – YouTube
It’s a sweet moment, and the way that the twins look at the balloons is so adorable.
Image: The Nowhere Twins – YouTube
The girls are excited to be surrounded by so many balloon clouds… even though they’re getting cold.
Image: The Now
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Dreadful, with adventure-mystery film production value and action sequences reminiscent of the (r)evolution Film Productions horror catalogue: Kane. Dreadful Film Downloadl and B18RB are able to detect phosphosites that are not predictive with other existing phosphoproteomics data. While an efficient method to predict potential phosphosites, even in the case of low abundance, requires a higher number of enrichment experiments, such experiments can now be reduced to one time-point as B18RB provides the means to precisely map phosphorylation sites and their stoichiometry on a particular substrate in response to a stimulus. B18RB is based on a two-step biotinylation procedure which requires neither pre-immunoprecipitation nor extensive washing steps as per existing strategies. We have tested the application of our strategy on a subset of RAW cells stimulated with LPS in order to produce a representative dataset for generating a positive/negative mapping experiment that is readily applicable for any substrate(s) of interest. Such a procedure is particularly useful in the case of proteins that are difficult to immunoprecipitate, as the resultant B-gel/lane provides access to a representative digest of the phosphoproteome for any given stimulus or cell type and enables the identification of protein phosphorylation events that are exclusively associated with a particular stimulus or cell type.
Methods {#Sec16}
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Cell culture {#Sec17}
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RAW264.7 mouse macrophage-like cell line (ATCC TIB-71) was grown in Dulbecco’s modified eagle medium (DMEM) (Thermo Fisher Scientific) supplemented with 10% (v/v) fetal calf serum (FCS) and 1% (v/v) penicillin/streptomycin. Cells were maintained at 37 °C, 95% relative humidity and 5% CO~2~.
Cell stimulation {#Sec18}
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Lipopolysaccharide from *Escherichia coli* (Sigma Aldrich) was used at a concentration of 1 µg/mL.
Biotinylation procedures {#Sec19}
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