Journal: PLoS ONE

Article Title: How Do Human Cells React to the Absence of Mitochondrial DNA?

doi: 10.1371/journal.pone.0005713

Figure Lengend Snippet: (A) Sub-organellar localization of Elac2 by western-blot analysis on mitochondria isolated from HeLa cells. Specificity of antibody reactivity was proved by the CRM obtained using an in vitro translated (i.v.) Elac2. A CRM was present in mitochondria (M), in the mitochondrial matrix (Ma) and membranes (Mb), and barely detectable in the nucleus (N). ETHE1 antibody was used as a mitochondrial matrix control protein. (B) Proteinase K protection assay by western-blot analysis on freshly isolated mitochondria from HeLa cells. Lane 1, cell lysate; lane 2, intact mitochondria; lane 3, mitochondria treated with 100 µg/ml of proteinase K; lane 4, mitochondria treated with 0.5% of Triton-X100. (C) Mitochondrial in vivo import of HA-tagged human Elac2 N-terminal polypeptide expressed in Cos-7 cells. Lane 1: marker 46 kDa (MW). lane 2: immunoprecipitation of total radiolabeled proteins. lane 3: immunoprecipitation of total radiolabeled proteins after the addition of valinomycin that determines the elimination of the mitochondrial ΔΨ: as a consequence, mitochondrial import is abolished. lane 4: immunoprecipitation of in vitro translation product (i.v.). Arrows indicate the Elac2 precursor (Elac2) and the mature form (mElac2). (D) Confocal immunofluorescence on Cos7 cells. The green fluorescence corresponds to Elac2HA-specific immunoreactivity. The red fluorescence corresponds to immunoreactivity specific to mtSSB. The two immunofluorescence patterns overlap, as shown by confocal merge. Magnification: 40×.

Article Snippet: An affinity purified polyclonal antibody against ELAC2 was from ProteinTech, while the polyclonal antibody against Ethe1 was raised in rabbit by Neosystem .

Techniques: Western Blot, Isolation, In Vitro, Control, In Vivo, Marker, Immunoprecipitation, Immunofluorescence, Fluorescence

Journal: DNA repair

Article Title: Release from quiescence stimulates the expression of human NEIL3 under the control of the Ras dependent ERK-MAP kinase pathway.

doi: 10.1016/j.dnarep.2012.01.007

Figure Lengend Snippet: Fig. 2. Human NEIL3 was highly expressed in cells that were in a proliferative state. HE, MRC-5, HaCaT and MCF-7 cells were serum starved at confluence and G0 released by replating 1:4 in serum containing culture. Normalized hNEIL3 relative transcript level (RQ) was measured at indicated time points after release by qPCR, using G0 HE cells as the

Article Snippet: The re-cleared extract was incubated with 1 g of NEIL3 PolyClonal ntibody (11621-1-AP, ProteinTech Group, Chicago, IL, USA) and 0 l Protein A/G PLUS Agarose (sc-2003, Santa Cruz Biotechnolgy).

Techniques:

Journal: DNA repair

Article Title: Release from quiescence stimulates the expression of human NEIL3 under the control of the Ras dependent ERK-MAP kinase pathway.

doi: 10.1016/j.dnarep.2012.01.007

Figure Lengend Snippet: Fig. 3. Human NEIL3 protein levels and DNA glycosylase activity mirrored hNEIL3 expression profile. Unsynchronized HaCaT cells were immunoprecipitated by NEIL3 antibody. (A) Western blot analysis of unsynchronized HaCaT cells that were immunoprecipitated with NEIL3 (+ NEIL3) or a control lacking NEIL3 antibody (− NEIL3). Arrows indicate the 70 kDa band sequenced, The IgG heavy chain and non-specific binding proteins. (B) Immunoprecipitated samples were incubated with a 32P-labeled single

Article Snippet: The re-cleared extract was incubated with 1 g of NEIL3 PolyClonal ntibody (11621-1-AP, ProteinTech Group, Chicago, IL, USA) and 0 l Protein A/G PLUS Agarose (sc-2003, Santa Cruz Biotechnolgy).

Techniques: Activity Assay, Expressing, Immunoprecipitation, Western Blot, Control, Binding Assay, Incubation, Labeling

Journal: DNA repair

Article Title: Release from quiescence stimulates the expression of human NEIL3 under the control of the Ras dependent ERK-MAP kinase pathway.

doi: 10.1016/j.dnarep.2012.01.007

Figure Lengend Snippet: Fig. 4. Human NEIL1 and NEIL3 were down- and up-regulated respectively upon release from quiescence. (A) HE fibroblasts, and (B) HaCaT cells were serum starved at confluence and G0 released by replating 1:4 in serum containing culture medium. Normalized hNEIL1, hNEIL2 and hNEIL3 relative transcript level (RQ) were measured at indicated time points after release by qPCR, using G0 cells as the reference sample for RQ calculation Experiment was repeated twice, one representative experiment is presented

Article Snippet: The re-cleared extract was incubated with 1 g of NEIL3 PolyClonal ntibody (11621-1-AP, ProteinTech Group, Chicago, IL, USA) and 0 l Protein A/G PLUS Agarose (sc-2003, Santa Cruz Biotechnolgy).

Techniques: