Journal: ACS chemical biology

Article Title: Human protamine-1 as an MRI reporter gene based on chemical exchange.

doi: 10.1021/cb400617q

Figure Lengend Snippet: Figure 2. Optimization of hPRM1. (a) Amino acid sequence alignment of protamine from salmon and humans. [*] indicates a conserved residue, [:] indicates groups with a strong similarity in their properties, and [.] indicates groups of weakly similar properties. (b) Partial, arginine-rich hPRM1 sequence of human- and E. coli-optimized genes. (c) Western blot of recombinant hPRM1 protein expressed in E. coli, using the human- or the E. coli-optimized DNA. (d) MTRasym plots and Western blot analysis (inset) of E. coli lysates from cells expressing either hPRM1 (red) or CD (blue). (e) The comparison of mean MTRasym values of hPRM1 (red bars) and CD (blue bars) at the 1.5 and 3.6 ppm frequency offsets. The mean MTRasym (+SD) were calculated from four independent measurements for each cell type (n = 4). CEST data were acquired at 11.7T, 37 °C, pH = 7.4, B1 = 4.7 μT, and tsat = 4000 ms.

Article Snippet: The gene encoding to hPRM1 (NM_002761) was obtained from Origene (Rockville, MD).

Techniques: Sequencing, Residue, Western Blot, Recombinant, Expressing, Comparison

Journal: ACS chemical biology

Article Title: Human protamine-1 as an MRI reporter gene based on chemical exchange.

doi: 10.1021/cb400617q

Figure Lengend Snippet: Figure 4. MR imaging of hPRM1 reporter gene expression in live cells in a three-dimensional cell culture system. (a) Bright-field microscopic images of encapsulated 293HEK cells. (b) T2-weighted images and overlaid MTRasym maps of encapsulated cells obtained at the 1.5 and 3.6 ppm frequency offsets. (c) p-values as a function of the frequency offset from water (Dw), as calculated using a Student’s t test (two- tailed distribution, paired test). Red line represents p-value of 0.05 (significance level). (d) Mean MTRasym (±SD) at 1.5 ppm calculated from three samples containing encapsulated cells (293hPRM1 and 293wt). CEST data were acquired at 11.7 T, 37 °C, pH = 7.4, B1 = 3.6 μT, and tsat = 3000 ms.

Article Snippet: The gene encoding to hPRM1 (NM_002761) was obtained from Origene (Rockville, MD).

Techniques: Imaging, Gene Expression, Cell Culture, Two Tailed Test

Journal: ACS chemical biology

Article Title: Human protamine-1 as an MRI reporter gene based on chemical exchange.

doi: 10.1021/cb400617q

Figure Lengend Snippet: Figure 3. CEST MRI of HEK293 cell extracts. (a) Western blot, using an anti-V5 antibody, showing hPRM1 expression. (b) MTRasym plots (±SD; n = 3). (c) Representative MTRasym maps obtained at the 1.5 and 3.6 ppm frequency offsets. (d) Mean MTRasym (±SD) at the 1.5 ppm and 3.6 ppm frequency offsets. CEST data were acquired at 11.7T, 37 °C, pH = 7.4, B1 = 4.7 μT, and tsat = 4000 ms.

Article Snippet: The gene encoding to hPRM1 (NM_002761) was obtained from Origene (Rockville, MD).

Techniques: Western Blot, Expressing

Journal: PLoS Genetics

Article Title: HENMT1 and piRNA Stability Are Required for Adult Male Germ Cell Transposon Repression and to Define the Spermatogenic Program in the Mouse

doi: 10.1371/journal.pgen.1005620

Figure Lengend Snippet: (A) A heatmap showing log2 fold change in differentially expressed genes in spermatocytes and their corresponding log2 fold change in round spermatids (round). Color coding represent log2 fold changes of genes. Prm1 , Tnp1 , Tnp2 , Prm2 indicate the position of transcripts subjected to further analysis. (B) A volcano plot showing expression changes of all detected genes in spermatocytes and round spermatids. Dots in cyan represent significantly differential expressed genes (FDR < 0.05) based on edgeR. Fold changes and p-values were calculated with edgeR. (C-G) qPCR of spermiogenic genes including (C) Tnp1 (D) Tnp2 (E) Prm1 (F) Prm2 (G) Gapdhs in 28 day-old Henmt1 WT/WT and Henmt1 PIN/PIN spermatocytes and round spermatids (n = 5 / genotype +/- SEM, * p<0.05, **p<0.01). White bar represents Henmt1 WT/WT and black bar is Henmt1 PIN/PIN . S’cytes = spermatocytes, S’tids = round spermatids. A two-tailed unpaired student T-test was performed for statistical analyses.

Article Snippet: Taqman Assays were purchased (Life Technologies) and used to quantitate the expression of Henmt1 isoform 1 and 2 (Mm00659237_m1), Ppi (Mm02342429_g1), Tnp1 (Mm04207755_g1), Tnp2 (Mm00726979_s1), Prm1 (Mm01342731_g1), Prm2 (Mm03048199_m1), Gapds (Mm00484668_m1), Spem1 (Mm01250806_g1), and Car2 (Mm00501576_m1).

Techniques: Expressing, Two Tailed Test

Journal: PLoS Genetics

Article Title: HENMT1 and piRNA Stability Are Required for Adult Male Germ Cell Transposon Repression and to Define the Spermatogenic Program in the Mouse

doi: 10.1371/journal.pgen.1005620

Figure Lengend Snippet: ChIP and qPCR analyses were performed on Henmt1 WT/WT and Henmt1 PIN/PIN spermatocytes (n = 5/genotypes, 3 biological replicates, * p<0.05, **p<0.01, *** p<0.001, **** p<0.0001). qPCR for the promoter regions of ( A ) Tnp1 , ( B ) Tnp2 , ( C ) Prm1 , ( D ) Prm2 , ( E ) Gapdhs , and ( F ) Ppia (as a house keeping control). Histone enrichment was normalised to histone H3. The data is presented in the ratio of Henmt1 PIN/PIN enrichment/ Henmt1 WT/WT enrichment. A two-tailed unpaired student T-test was performed for statistical analyses.

Article Snippet: Taqman Assays were purchased (Life Technologies) and used to quantitate the expression of Henmt1 isoform 1 and 2 (Mm00659237_m1), Ppi (Mm02342429_g1), Tnp1 (Mm04207755_g1), Tnp2 (Mm00726979_s1), Prm1 (Mm01342731_g1), Prm2 (Mm03048199_m1), Gapds (Mm00484668_m1), Spem1 (Mm01250806_g1), and Car2 (Mm00501576_m1).

Techniques: Control, Two Tailed Test

Journal: Scientific Reports

Article Title: CRISPR/Cas9-mediated knockout of Mct8 reveals a functional involvement of Mct8 in testis and sperm development in a rat

doi: 10.1038/s41598-020-67594-2

Figure Lengend Snippet:

Article Snippet: Prm1 , Rn02345725_g1.

Techniques: Gene Expression