Journal:

Article Title: In Vivo Rescue of a Silent tax -Deficient Bovine Leukemia Virus from a Tumor-Derived Ovine B-Cell Line by Recombination with a Retrovirally Transduced Wild-Type tax Gene

doi:

Figure Lengend Snippet: Detection of LTaxSN and BLV sequences in YR2 and YR2LTaxSN cells. (A) Southern blot analysis of SacI-digested genomic DNA. Twenty micrograms of digested DNA prepared from YR2 (lanes 1, 3, 5, and 7) or YR2LTaxSN (lanes 2, 4, 6, and 8) cells was electrophoresed through a 0.8% agarose gel, transferred to nylon membranes, and hybridized with 32P-labelled ENV, NEO, MoMLV LTR, and TAX probes. (B) PCR amplification of LTaxSN and BLV sequences. Genomic DNA isolated from parental YR2 (lanes 1, 5, 9, 13, and 17) and Rat-2 (lanes 4, 8, 12, 16, and 20) cells and from YR2LTaxSN (lanes 2, 6, 10, 14, and 18) and Rat-2LTaxSN (lanes 3, 7, 11, 15, and 19) cells was amplified by PCR with LTaxSN-specific primers P1/T2, P1/P2, T1/P2, and N1/N2 or BLV-specific primers T6/U3 and analyzed by Southern blot hybridization with 32P-labelled TAX and NEO probes.

Article Snippet: FLK (a BLV-infected fetal lamb kidney cell line), Rat-2 (a control cell line, ATCC CRL1764), and Rat-2 derivatives (Rat-2 LTaxSN and Rat-2 NUNL ) were grown in Dulbecco’s modified Eagle’s medium supplemented with 10% FCS, 1 mM sodium pyruvate, 2 mM glutamine, nonessential amino acids, and 100 μg of kanamycin per ml.

Techniques: Southern Blot, Agarose Gel Electrophoresis, Amplification, Isolation, Hybridization

Journal:

Article Title: In Vivo Rescue of a Silent tax -Deficient Bovine Leukemia Virus from a Tumor-Derived Ovine B-Cell Line by Recombination with a Retrovirally Transduced Wild-Type tax Gene

doi:

Figure Lengend Snippet: Detection of BLV and LTaxSN transcripts in YR2LTaxSN cells. (A) Ten micrograms of total RNA isolated from YR2, YR2LTaxSN, Rat-2, Rat-2LTaxSN, or FLK cells was analyzed by Northern blot hybridization with 32P-labelled TAX (left panel) and NEO (right panel) probes. Sizes of mRNAs are indicated. (B) RT-PCR detection of the BLV 2.1-kb doubly spliced transcript in total RNA from YR2 (lanes 1 and 2), YR2NUNL (lanes 3 and 4), Rat-2LTaxSN (lanes 5 and 6), YR2LTaxSN (lanes 7 and 8), and FLK (lanes 9 and 10) cells with two sets of splice-specific primers, EA/C3 (lanes 1, 3, 5, 7, and 9) and EA/T2 (lanes 2, 4, 6, 8, and 10). Amplification products were electrophoresed through a 1% agarose gel, transferred to nylon membranes, and hybridized with a 32P-labelled total BLV probe. Positions and molecular sizes of amplified products are indicated.

Article Snippet: FLK (a BLV-infected fetal lamb kidney cell line), Rat-2 (a control cell line, ATCC CRL1764), and Rat-2 derivatives (Rat-2 LTaxSN and Rat-2 NUNL ) were grown in Dulbecco’s modified Eagle’s medium supplemented with 10% FCS, 1 mM sodium pyruvate, 2 mM glutamine, nonessential amino acids, and 100 μg of kanamycin per ml.

Techniques: Isolation, Northern Blot, Hybridization, Reverse Transcription Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis

Journal: bioRxiv

Article Title: mTORC1 signaling is not essential for the maintenance of muscle mass and function in adult sedentary mice

doi: 10.1101/738294

Figure Lengend Snippet: iRAmKO mice have a reduced oxidative capacity and shift the fiber-type composition towards slower fiber types. (A) The activity of oxidative enzymes was examined by NADH-TR staining on TA cross-sections of 3-month iRAmKOs. Scale bar = 50 µm. (B) Relative mRNA levels of Ppargc1α (PGC1α), Mb (myoglobin), Cycs (cytochrome c) and Cox5b (cytochrome c oxidase subunit 5B) in the soleus muscle of 3-month iRAmKOs. (C) Immunohistochemical (IHC) staining against laminin (white), MyHC type 2B (cyan), MyHC type 2A (red), MyHC type 1/slow (yellow) and type 2X (unstained). The top image is the TA and the lower image is the soleus of a 5-month iRAmKO and control. Scale bar = 200 µm. (D) and (E) Quantification of the fiber type composition in TA (D) and soleus (E) of 5-month iRAmKOs and their control littermates. n ≥ 3 for 3-month iRAmKOs and n = 4 for 5-month iRAmKOs. Values represent the mean ± SEM. Significance was assessed using two-tailed unpaired student’s t -test: *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The primary antibodies against MyHC 2b (1:100; #BF-F3; DSHB, Iowa, USA), MyHC 2a (1:200; #SC-71; DSHB), MyHC 1 (1:50; BA-D5; DSHB) and laminin (1:160; #ab11575; Abcam) were diluted in PBS containing 10% goat serum (Biological industries) and added for 1 h at RT.

Techniques: Activity Assay, Staining, Immunohistochemical staining, Immunohistochemistry, Control, Two Tailed Test