Effects of bafilomycin A1 pretreatment on transgene expression.

<p> COS7 and HEK293 cells were treated with either buffer in the control (Ctrl) or an endocytic trafficking inhibitor, bafilomycin A1 (Baf A1), at 1 μM for 1 hour prior to electrotransfection; and the eTE was quantified at 24 hours post electrotransfection. (n = 4, *P<0.05, Mann-Whitney U test).</p

Release of rhodamine B-labeled dextran from endosomes following light treatment.

<p>Intracellular vesicles were preloaded with the photosensitizer. (a) The image of a COS7 cell before light treatment. It was taken at 20 min after the cell was incubated at 37°C with dextran (10,000 MW). The light treatment was performed immediately after the image acquisition. (b) The image of the same cell at 10 min after light treatment. The dextran was punctate within endosomes before light treatment (top panel). However, upon light treatment, dextran diffused out of endosomes, and spread in the cytosol (bottom panel).</p

Efficiency of cell transfection with Poly-L-lysine (PLL).

<p>Intracellular vesicles were preloaded with the photosensitizer. COS7 cells were transfected with PLL-pDNA complexes for 4 hours at 37°C. Then, the cells were exposed to blue light for 0 (i.e., no light (NL) control), 1 min, and 5 min. The transfection efficiency (TE) was quantified at 24 hours post transfection. (n = 4, *P<0.05, Mann-Whitney U test).</p

Effects of lysosomotropic agents pretreatment on transgene expression.

<p> COS7 and HEK293 cells were treated with either buffer in the control (Ctrl) or a lysosomotropic agent for 4 hours prior to electrotransfection: (a) chloroquine (CQ) at 100 μM, and (b) ammonium chloride (AC) at 10 mM. The treatments had statistically insignificant effects on eTE quantified at 24 hours post electrotransfection. (n = 4, P > 0.05, Mann-Whitney U test).</p

Additional file 5: Figure S1. of Long noncoding RNA TUG1 is downregulated in non-small cell lung cancer and can regulate CELF1 on binding to PRC2

Digestion and PCR amplification of the TUG1-4C procedure. (A) Agarose gel (0.8 %, wt/vol) of the undigested (left lane) and primary digested (right lane) sample (HindIII, six-cutter). (B) Agarose gel (2.0 %, wt/vol) of the secondary digested sample (CViQI, four-cutter). (C) Inverse PCR amplification of the 4C samples generated amplified sequences of a wide range of sizes. (JPG 26 kb

Additional file 2: of Long noncoding RNA TUG1 is downregulated in non-small cell lung cancer and can regulate CELF1 on binding to PRC2

RIP and DNA ChIP. (DOCX 15 kb

Carbodicarbenes: Unexpected π‑Accepting Ability during Reactivity with Small Molecules

An investigation of carbodicarbenes, the less explored member of the carbenic complex/ligand family has yielded unexpected electronic features and concomitant reactivity. Observed 1,2-addition of E–H bonds (E = B, C, Si) across the carbone central carbon and that of the flanking N-heterocyclic carbene (NHC) fragment, combined with single-crystal X-ray studies of a model Pd complex strongly suggests a significant level of π-accepting ability at the central carbon of the NHC moiety. This feature is atypical of classic NHCs, which are strong σ-donors, with only nominal π-accepting ability. The unanticipated π-acidity is critical for engendering carbodicarbenes with reactivity more commonly observed with frustrated Lewis pairs (FLPs) rather than the more closely related NHCs and cyclic (alkyl)­(amino)­carbenes (CAACs)