Propofol attenuates CoCl₂-induced TNF-α generation.

<p>A, In BV2 cells, 500 μM CoCl₂ treatment induced TNF-α generation in a time-dependent manner, and 8h treatment significantly increased TNF-α generation. B, propofol attenuated CoCl₂-induced TNF-α generation in a suitable concentration. 25μM propofol significantly reduced TNF-α generation. C, propofol alone had no effect on TNF-α generation. (* p < 0.05 vs. control, # p < 0.05 vs. CoCl₂ treatement, n = 5, Data were shown as mean ± SD).</p

Effects of propofol on CoCl₂ induced the destruction of intracellular Ca²⁺ homeostasis and the phosphorylation of CAMKIIα, ERK, NF-κB.

<p>A and B, CoCl₂-increased intracellular Ca²⁺ concentration was attenuated by propofol. C, D, E, F, G and H, CoCl₂-induced the phosphorylation of CAMKIIα, ERK and NF-κB were attenuated by propofol. (* p < 0.05 vs. control, # p < 0.05 vs. CoCl₂ treatement, n = 5, Data were shown as mean ± SD).</p

Effects of propofol, BAPTA-AM, KN93, U0126 on CoCl₂ induced the phosphorylation of CAMKIIα, ERK, NF-κB.

<p>CoCl₂-induced the phosphorylation of CAMKIIα, ERK and NF-κB were attenuated by propofol, BAPTA-AM, KN93 and U0126. (* p < 0.05 vs. control, # p < 0.05 vs. CoCl₂ treatement, n = 5, Data were shown as mean ± SD).</p

Effects of propofol, BAPTA-AM, KN93, U0126, INCB039110 on CoCl₂ induced TNF-α generation.

<p>CoCl₂ treatment induced TNF-α generation was attenuated by propofol and BAPTA-AM, KN93, U0126. The effect of propofol on TNF-α generation was reversed by INCB039110. (* p < 0.05 vs. control, # p < 0.05 vs. CoCl₂ treatement, n = 5, Data were shown as mean ± SD).</p

Effects of propofol, BAPTA-AM, KN93, U0126 on CoCl₂ induced cell apoptosis.

<p>A and B, CoCl₂-increased cell apoptosis was attenuated by propofol, BAPTA-AM, KN93 and U0126. C and E, CoCl₂-induced cleaved caspase 3 overexpression was attenuated by propofol, BAPTA-AM, KN93 and U0126. D and F, CoCl₂, propofol, BAPTA-AM, KN93 and U0126 had no effect on the expression of pro-caspase 3. (* p < 0.05 vs. control, # p < 0.05 vs. CoCl₂ treatement, n = 5, Data were shown as mean ± SD).</p

EF1a-GFP targeting in the <i>pRosa26</i> locus.

<p>(A) Schematic representive of the EF1a-GFP targeting vector and a segment of the <i>pRosa26</i> locus. SA, splice acceptor. The blue dashed line indicates the band size (1.8 kb) in the 5′ arm PCR analysis using F and R primer set. (B) 5′ arm PCR analysis of targeted cell clone. M, DNA marker. P6, PPNT6 plasmid. WT, wild type DNA. (C) GFP expression in correctly targeted cells (200×). (D) GFP expression in cloned blastocysts from targeted cells (100×). (E) DNA methylation status of EF1a in <i>pRosa26</i> targeted cells and cloned blastocysts. The methylation status was detected by the bisulfite sequencing. Methylated and non-methylated CpG dinucleotides of each clone are illustrated with closed and open circles, respectively.</p

Identification of the <i>pRosa26</i> locus.

<p>(A) A diagram of the <i>pRosa26</i> locus on chromosome 13. (B) Expression of <i>pRosa26</i> ncRNA in various tissues relative to b-actin by Q-PCR. Error bars are mean±SD.</p

MyoP-Fst targeting in the <i>pRosa26</i> locus.

<p>(A) Schematic representive of the MyoP-Fst targeting vector and a segment of the <i>pRosa26</i> locus. SA, splice acceptor. The red and black dashed lines indicate the knockin (1.2 kb) and WT (3.0 kb) band sizes expected after XhoI/XbaI digestion in the Southern blot. The yellow dashed line indicates the Fst band size (1.85 kb) expected after HindIII/BamHI digestion in the Southern blot. The blue dashed line indicates the band size (1.8 kb) in the 5′ arm PCR analysis using F and R primer set. (B) Southern blot of cloned pigs. 5 correctly targeted pigs were confirmed. (C) Fst expression in ear of 3 Fst-KI pigs by Western blot. (D) Fst expression relative to b-actin in various tissues of Fst-KI pig by Q-PCR. (E) Fst expression in various tissues of Fst-KI pig by Western blot. (F) DNA methylation status of MyoP in various tissues of Fst-KI pig. (G) Fst-KI pigs. 130731-1 and -2 are marked with *. WT, wild type.</p

The development of GFP-KI cloned embryos <i>in</i><i>vitro.</i>

<p>Note: Different superscripts mean significant difference (p<0.05).</p><p>The development of GFP-KI cloned embryos <i>in</i><i>vitro.</i></p

The development of Fst-KI cloned embryos <i>in</i><i>vivo.</i>

<p>Note: 130731-1 and 130731-2 pigs are still alive.</p><p>The development of Fst-KI cloned embryos <i>in</i><i>vivo.</i></p