Journal: PLoS ONE

Article Title: Streptococcus iniae M-Like Protein Contributes to Virulence in Fish and Is a Target for Live Attenuated Vaccine Development

doi: 10.1371/journal.pone.0002824

Figure Lengend Snippet: (A) Adherence and (B) invasion characteristics of WT K288, the isogenic Δ simA allelic mutant, and the naturally M-deficient WT 02161A S. iniae strain for the fish epithelial cell line WBE27. (C) Survival of WT K288, WT 02161A, and the Δ simA mutant upon co-incubation with CLC fish macrophage/monocytes for 2, 4, or 18 h. Significance indicated as: * P <0.05, ** P <0.005, *** P <0.0005. Data are presented as mean±SEM from two-tailed t-tests.

Article Snippet: The adherent CLC carp monocytic/macrophage cell line (European Collection of Cell Cultures no. 95070628) and the WBE27 white bass embryonic epithelial cell line (ATCC no. CRL-2773) were grown at 28°C with 5% CO 2 .

Techniques: Mutagenesis, Incubation, Two Tailed Test

Journal: Protein & Cell

Article Title: The chemical reprogramming of unipotent adult germ cells towards authentic pluripotency and de novo establishment of imprinting

doi: 10.1093/procel/pwac044

Figure Lengend Snippet: 5C-gPSCs had true pluripotency identified by tetraploid complementation assay. (A) 5C-gPSC 4N-comp pups and their corresponding placentae. Note: * Pups survived beyond 4 days postnatal; # Pup died within 4 days postnatal. (B) Simple sequence length polymorphism (SSLP) analysis for lineage identification covers markers from different chromosomes, and the 4N-comp mice showed a polymorphic pattern similar with that from the F1-5 ESC, 5C-gPSC-S4, or gPSC-S2 cells originating from B6D2F1 mice, and different from the ICR, C57, or DBA mice. M denotes molecular mass marker. (C) Genotyping analysis of Oct4 -EGFP from F1-5 ESC, 5C-gPSC-S4, or gPSC-S2 4N-comp mice. (D) Bright field (left), fluorescence (middle) and merged (right) images of E3.5 Oct4 -EGFP blastocysts flushed from an ICR female mouse mated with an 5C-gPSC-S4 4N-comp mouse. Scale bar, 100 μm. (E) An 11-week-old 5C-gPSC-S4 4N-comp mouse with a uniformly black coat and its F 1 progeny from its mating to an ICR female mouse. (F) Bar plot showing the developmental rate of full-term 4N-comp pups derived from PSCs, numbers of animals obtained per total number of transferred embryos are shown. Groups are indicated by colors. See for details. (G) Dot plot showing the body weight of 4N-comp mice born by Caesarean section in this study on day 0 postnatal. Groups are indicated by colors. Unpaired two-tailed Student’s t -test, * P < 0.05, ns, not significant. (H) Survival curves of 4N-comp mice obtained in this study. n, total numbers of 4N-comp mice derived from each group. Groups are indicated by colors. (I) Growth curves of 4N-comp mice obtained in this study at the indicated weeks postnatal. Groups are indicated by colors. (J) Bar plot showing the DNA methylation level of the whole genome (left) and total ICRs (right) in ESCs (p10), 5C-gPSCs (p25), gPSCs (p25), respectively. Unpaired two-tailed Student’s t -test, P values were shown above the lines, ** P < 0.01, ns, not significant. (K) Heatmaps showing the mean methylation levels of the paternal ICRs in SSCs (p20), gPSCs (p25), 5C-gPSCs (p25), ESCs (p10). The color key from dark blue to red indicates low to high levels, respectively. (L) Line plot shows the mean methylation levels of Dlk1-Dio3 region in SSCs (p20), 5C-gPSCs (p25), ESCs (p10), gPSCs (p25), respectively. Grey shadows indicate gene regions, the red shadow indicates IG- Gtl2 ICR region. (M) Box plots show the mean DNA methylation levels across gene bodies (from transcription start site (TSS) to transcription end site (TES)) and the 50-bp flanking regions of each miRNA encoded locus in Dlk1-Dio3 region in ESCs, gPSCs and 5C-gPSCs. One dot represents an miRNA encoded locus, the same loci in distinct PSCs are connected by grey lines. Paired two-tailed Student’s t -test was performed between ESCs and other PSCs. (N) Heatmaps showing the mean methylation levels of the maternal ICRs in SSCs (p20), gPSCs (p25), 5C-gPSCs (p25), ESCs (p10). The color key from dark blue to red indicates low to high levels, respectively. (O) qPCR analysis of relative expression of Igf2r , Kcnq1ot1 , Peg3 , and Snrpn normalized to the geometric mean of Rps2 and Gapdh in ESCs, gPSCs, and 5C-gPSCs. Unpaired two-tailed Student’s t -test, ** P < 0.01, *** P < 0.001, ns, not significant. See also .

Article Snippet: For tetraploid embryo complementation, two-cell stage embryos were collected from oviducts of ICR females (white coat color) and electrofused by Electro Cell Manipulator (BTX, EMC2001) to produce one-cell tetraploid embryos that were then cultured in M2 medium (Sigma-Aldrich).

Techniques: Sequencing, Marker, Fluorescence, Derivative Assay, Two Tailed Test, DNA Methylation Assay, Methylation, Expressing