Engineering of polyketide synthases: How close are we to the reality?

<p>(A) Oocytes and parthenogenetic embryos were immunostained with the anti-H3K36me3 antibody, which was then localized with a FITC-conjugated secondary antibody (green). DNA was stained with DAPI (blue). NSN and SN are two types of GV stage oocytes. MI, metaphase I stage oocytes; MII, metaphase II stage oocytes. Scale bar = 20 µm. (B) Relative intensities of fluorescence signals from H3K36me3. Total 23 oocytes and 47 embryos were analyzed in triplicate in this experiment. Bars represent least-squares showed the standard error in each group. P-values <0.05 were considered statistically significant. ^a,b,c Values represent least-squares with different superscripts are significantly different (P<0.05).</p

Changes in H3K36me3 status in porcine IVF and SCNT embryos.

<p>(A) Sperm, MII stage oocytes coincubated with sperm in mTBM medium for 1 h (IVF 1 h), and IVF embryos cultured in PZM-3 medium for 18 h (IVF) were immunostained with the anti-H3K36me3 antibody. (B) A single fetal fibroblast cell injected into the perivitelline space of a denuded oocyte without activation (Embedded), a nuclear-transferred embryo cultured in PZM-3 for 1 h post activation (1h), a nuclear-transferred embryo cultured in PZM-3 for 18 h after activation (18h), and other stages of SCNT embryos were immunostained with the anti-H3K36me3 antibody. The antibody was localized with a FITC-conjugated secondary antibody (green). DNA was stained with DAPI (blue). The arrowhead indicates sperm and the arrow indicates the nuclei of donor cell. Scale bar = 20 µm. (C) Relative intensities of fluorescence signals from H3K36me3. Total 41 embryos were analyzed in triplicate in this experiment. Bars represent least-squares showed the standard error in each group. P-values <0.05 were considered statistically significant. ^a,b,c Values represent least-squares with different superscripts are significantly different (P<0.05).</p

H3K36 methylation status and association with transcriptional activity in porcine fetal fibroblasts.

<p>(A) Cells were cultured in medium containing 2.5 mM FU for 30 min, fixed, and immunostained with antibodies against FU and H3K36me1, -me2 or -me3. Primary antibodies were detected with FITC-conjugated (green) and TR-conjugated (red) secondary antibodies. DNA was stained with DAPI (blue). Enlargements (bottom insets) were the area pointed by thick arrows in merge insets showing transcriptional active sites (labeled with FU), H3K36 methylations and their overlap. The asterisk indicates the nucleolus. (B) The overall overlap rates between H3K36 methylations and transcriptional active sites were analyzed using total 30 dots indicated by the thin and thick arrows in (A).</p

Effect of flavopiridol treatment on H3K36 methylation status in porcine fetal fibroblasts.

<p>Cells were treated with 200(FP) for 30 min, followed by combined treatment with 200 nM flavopiridol and 2.5 mM FU for an additional 30 min. Cells were then fixed and immunostained with antibodies against FU and H3K36me1, -me2, or -me3. Primary antibodies were detected with FITC-conjugated (green) and TR-conjugated (red) secondary antibodies. DNA was stained with DAPI (blue).</p

The different sizes of clear corneal incisions for phacoemulsification in eyes with the different RK incisions.

<p>(A) The 3.2mm clear corneal incisions in eye with 8 incisions. (B) The 3.2mm clear corneal incisions in eye with 12 incisions. (C) The 3.2mm clear corneal incisions in eye with 16 incisions. (D) The 3.0mm clear corneal incisions in eye with 8 incisions. (E) The 2.2mm clear corneal incisions in eye with 12 incisions. (F) The 2.0mm clear corneal incisions in eye with 16 incisions.</p

The result of comparing to pre- and post-operative vision, astigmatism, and corneal endothelial cell density of 30 eyes.

<p>The result of comparing to pre- and post-operative vision, astigmatism, and corneal endothelial cell density of 30 eyes.</p

Two eyes with intraoperative RK incision dehiscence.

<p>(A) RK wound dehiscence occurred during the process of phacoemulsification. (B) RK wound dehiscence occurred during the process of perfusion aspiration.</p

The results of the spherical equivalent and the error between postoperative spherical equivalent and the calculated target refraction by the SRK-T formula and HOFFER-Q modified formula.

<p>The results of the spherical equivalent and the error between postoperative spherical equivalent and the calculated target refraction by the SRK-T formula and HOFFER-Q modified formula.</p

Space between two neighboring RK incisions at the cornea limbus (mm).

<p>Space between two neighboring RK incisions at the cornea limbus (mm).</p

The independent sample t test of pre- and post-operative vision, astigmatism, and corneal endothelial cell density stratified by different corneal incision size in 8 cuts group.

<p>The independent sample t test of pre- and post-operative vision, astigmatism, and corneal endothelial cell density stratified by different corneal incision size in 8 cuts group.</p