Journal: BMC Research Notes

Article Title: Cloning of NruI and Sbo13I restriction and modification sstems in E. coli and amino acid sequence comparison of M.NruI and M.Sbo13I with other amino-methyltransferases

doi: 10.1186/1756-0500-3-139

Figure Lengend Snippet: Conserved amino acid sequence blocks in the multiple sequence alignment of M.NruI and M.Sbo13I family sequences . The homologous sequences found in GenBank are: gi_148658155 (YP_001278360), gi_149918461 (ZP_01906951), gi_225022629 (ZP_03711821), gi_252123721 (ZP_04834998) and putative methyltransferase M.Rca13941_ORF2728P. The consensus amino acid residues are indicated below the 7 actual sequences. The h and e indicate secondary structural prediction for α-helix (indicated in red) and β-strand (indicated in blue) respectively. Conserved amino acid motifs (blocks) I through X are shown above the amino acid sequences.

Article Snippet: The GenBank protein and gene IDs are described as following: ZP_01906951 (gi_149918461, PPSIR1_36012 from Plesiocystis pacifica SIR-1), ZP_03711821 (gi_225022629, CORMATOL_02672 from Corynebacterium matruchotii ), ZP_04834998 (gi_252123721, M.Cma ORF1763, ATCC_14266 from Corynebacterium matruchotii ), and M.Rca13941ORF2728P (DSM 13941 from Roseiflexus castenholzii ).

Techniques: Sequencing

Journal: Oncotarget

Article Title: Cardiac glycoside bufalin blocks cancer cell growth by inhibition of Aurora A and Aurora B activation via PI3K-Akt pathway

doi: 10.18632/oncotarget.24475

Figure Lengend Snippet: ( A ) Thymidine synchronized HT-29 cells were released for 9 h in the presence of vehicle (0.1% DMSO), bufalin (100 nM), LY249002 (LY; 20 μM), rapamycin (Rap; 200 nM), PD98059 (PD; 20 μM), or NAC (10 mM). The phospho-Aurora A (Thr288)/Aurora B (Thr232) and β-actin were analyzed by Western blot. ( B ) HT-29 cells were transfected with Erk1/2 siRNA for 33 h, exposed to 2 mM thymidine for 15 h and followed by harvest after 9 h of release into 10% FBS medium. The protein levels of p-Aurora A/B were analyzed by Western blot. ( C – D ) HT-29 cells were transfected with siRNAs against sodium pump α1 and α3 for 33 h, exposed to 2 mM thymidine for 15 h and followed by harvest after 9 h of release into 10% FBS medium. (C) The knockdown efficiency of α1 siRNA and α3 siRNA was analyzed by Western blot. (D) Effect of sodium pump on the activities of PI3K, Akt, mTOR, and Aurora A/B was analyzed by Western blot. ( E – F ) Before entering into mitosis, thymidine-synchronized HT-29 cells were released for 5 h and then exposed to bufalin (100 nM), LY294002 (LY; 100 μM), a combination of two drugs, or equal amount of DMSO for 4 h. (E) Effect of PI3K inhibitor LY294002 on active form of Akt. (F) Effect of PI3K pathway on cell progression. ( G – H ) HT-29 cells were transfected with empty vector or HA-Akt for 24 h and then treated with bufalin for 24 h, or cotransfected with HA-Akt plus α3 siRNA for 48 h. (H) Cell number was counted for each group. (G) The protein levels of HA-Akt, p-Akt, p-Aurora A (Thr288)/B (Thr232), Plk1 and β-actin were analyzed by Western blot. ( I ) Before entering into mitosis, thymidine-synchronized HT-29 cells were released for 5 h and then exposed to bufalin (100 nM), LY294002 (LY; 100 μM), a combination of two drugs, or equal amount of DMSO for 4 h. The protein levels of NF-κB p65, HIF1α and Plk1 were analyzed by Western blot. ( J ) HT-29 cells were transfected with Plk1 siRNA for 33 h, exposed to 2 mM thymidine for 15 h and followed by harvest after 9 h of release into 10% FBS medium.

Article Snippet: The α3 siRNA (sc-36012) for α3 isoform of sodium pump was obtained from Santa Cruz.

Techniques: Western Blot, Transfection, Plasmid Preparation

Journal: Upsala Journal of Medical Sciences

Article Title: Expression of Na/K-ATPase subunits in the human cochlea: a confocal and super-resolution microscopy study with special reference to auditory nerve excitation and cochlear implantation

doi: 10.1080/03009734.2019.1653408

Figure Lengend Snippet: Antibodies used in the present investigation.

Article Snippet: Na/K-ATPase (α3) , polyclonal , 1:800 , goat , sc-16052 , Santa Cruz Biotechnology.

Techniques:

Journal: Upsala Journal of Medical Sciences

Article Title: Expression of Na/K-ATPase subunits in the human cochlea: a confocal and super-resolution microscopy study with special reference to auditory nerve excitation and cochlear implantation

doi: 10.1080/03009734.2019.1653408

Figure Lengend Snippet: Expression of Na/K-ATPase subtypes.

Article Snippet: Na/K-ATPase (α3) , polyclonal , 1:800 , goat , sc-16052 , Santa Cruz Biotechnology.

Techniques: Expressing

Journal: Upsala Journal of Medical Sciences

Article Title: Expression of Na/K-ATPase subunits in the human cochlea: a confocal and super-resolution microscopy study with special reference to auditory nerve excitation and cochlear implantation

doi: 10.1080/03009734.2019.1653408

Figure Lengend Snippet: Confocal microscopy and composite micrograph showing the expression of Na/K-ATPase β1 in a mid-modiolar sections of an adult human cochlea. The interrupted line shows position of Reissner’s membrane. There is a strong expression of Na/K-ATPase at the spiral prominence, stria vascularis, and the plasma membranes of the spiral ganglion cell bodies. The ganglion cells express connexin30 (insets). Framed area in upper inset is shown in higher magnification in lower left inset. Cochlear turns are denoted by 1–3. Fibrocyte types in the lateral wall are marked I–V. Framed areas are magnified in and Supplementary Figure 2C (available online).

Article Snippet: Na/K-ATPase (α3) , polyclonal , 1:800 , goat , sc-16052 , Santa Cruz Biotechnology.

Techniques: Confocal Microscopy, Expressing

Journal: Upsala Journal of Medical Sciences

Article Title: Expression of Na/K-ATPase subunits in the human cochlea: a confocal and super-resolution microscopy study with special reference to auditory nerve excitation and cochlear implantation

doi: 10.1080/03009734.2019.1653408

Figure Lengend Snippet: A: SR-SIM showing Na/K-ATPase β1 expression (red) in the type I cell soma plasma membranes. B: Framed area in A shows the expression of Na/K-ATPase β1 at the axon hillock in higher magnification. This isoform is not expressed in the cell membrane of the satellite glial cells (inset). C: CM of the spiral ganglion framed in . Framed area is magnified in D. Some cells lie juxtaposed (*) and show stronger expression of Na/K-ATPase, which is also verified in the false color display in E. F: A type I spiral ganglion cell is surrounded by a satellite glial cell that shows strong expression of Na/K-ATPase α1 (red). The nerve cell body expresses Nav1.6 (green).

Article Snippet: Na/K-ATPase (α3) , polyclonal , 1:800 , goat , sc-16052 , Santa Cruz Biotechnology.

Techniques: Expressing

Journal: Upsala Journal of Medical Sciences

Article Title: Expression of Na/K-ATPase subunits in the human cochlea: a confocal and super-resolution microscopy study with special reference to auditory nerve excitation and cochlear implantation

doi: 10.1080/03009734.2019.1653408

Figure Lengend Snippet: A: Confocal microscopy of Na/K-ATPase β1 expression in neurons of the human organ of Corti. The basolateral membranes of Hensen cells (HCs) are positive as well. The Corti tunnel is partly collapsed. Neurons beneath the IHC, outer spiral bundle (OSB), and tunnel spiral bundle show intense expression of the β1 isoform. B: Scanning electron microscopy of an OSB beneath the OHC. Framed area is magnified in C. Inset in D shows β1 expression of the axons. The bundle is believed to contain both efferents and afferents. D and E: Scanning electron microscopy of pre-terminal fibers (yellow) at the habenula perforata. Fiber diameter is between 0.5 and 1.0 μm.

Article Snippet: Na/K-ATPase (α3) , polyclonal , 1:800 , goat , sc-16052 , Santa Cruz Biotechnology.

Techniques: Confocal Microscopy, Expressing, Electron Microscopy

Journal: Upsala Journal of Medical Sciences

Article Title: Expression of Na/K-ATPase subunits in the human cochlea: a confocal and super-resolution microscopy study with special reference to auditory nerve excitation and cochlear implantation

doi: 10.1080/03009734.2019.1653408

Figure Lengend Snippet: (a) A–C: Na/K-ATPase β1 expression in different turns of the human cochlea. Intensity differences are represented by false color display. There is highest intensity in type II fibrocytes beneath the spiral prominence epithelium. Hensen cells are positive (arrow in A), There is no expression of Na/K-ATPase in the Reissner’s membrane. D–F: Na/K-ATPase β1 expression in the spiral ganglion neurons at different turns showing some variation. Some cells are dislodged basally (arrow in D). Framed area in E is magnified in b) Higher magnification of Na/K-ATPase β1 expression that was revealed in . Framed area is magnified in the inset. Note lack of β1 expression in Reissner’s membrane, intermediate cells (IC), basal cells (BCs), and types I and III fibrocytes. Upper and lower limits of the stria vascularis (marked with asterisks) also lack β1 expression. Inset shows strong Na/K-ATPase activity around the IHC representing neurons and phalangeal and inner sulcus cells. Interdental cells, spiral limbus fibrocytes, and Hensen cells show moderate intensity. There is high activity in the type II fibrocytes of the lateral wall.

Article Snippet: Na/K-ATPase (α3) , polyclonal , 1:800 , goat , sc-16052 , Santa Cruz Biotechnology.

Techniques: Expressing, Activity Assay

Journal: Upsala Journal of Medical Sciences

Article Title: Expression of Na/K-ATPase subunits in the human cochlea: a confocal and super-resolution microscopy study with special reference to auditory nerve excitation and cochlear implantation

doi: 10.1080/03009734.2019.1653408

Figure Lengend Snippet: Immunohistochemistry of the human cochlea in a temporal bone obtained at autopsy. A: There is a strong expression of Na/K-ATPase α3 subunit in the spiral ganglion cell bodies, membrane, and nerve fibers. Both type I and type II cells (D) are positive. B: There is a high expression of α3 in the spiral lamina fibers and neurons beneath the IHCs and OHCs. Basal tunnel fibers (BTF-afferents) (inset in B) and efferent tunnel-crossing fibers (TCFs) (C) also express the α3 subunit. E: α1 is intensively expressed in the satellite glial cells at the axon hillock region using CM F: Corresponding type I cell shows α3 expression in the neuron at the axon hillock. OSB: outer spiral bundle; TM: tectorial membrane.

Article Snippet: Na/K-ATPase (α3) , polyclonal , 1:800 , goat , sc-16052 , Santa Cruz Biotechnology.

Techniques: Immunohistochemistry, Expressing