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1.
FIG. 3.

FIG. 3. From: Human Primordial Germ Cell Formation Is Diminished by Exposure to Environmental Toxicants Acting through the AHR Signaling Pathway.

Western analysis of AHR in 293FT and hESCs. Upper panels are Western blots against AHR in 293FT cells and hESCs, and lower panels are Western blots of GAPDH as loading controls of the same samples. Asterisk indicates second band, which may be a posttranslationally modified form of AHR in 293FT cells.

Kehkooi Kee, et al. Toxicol Sci. 2010 September;117(1):218-224.
2.
FIG. 4.

FIG. 4. From: Human Primordial Germ Cell Formation Is Diminished by Exposure to Environmental Toxicants Acting through the AHR Signaling Pathway.

Gene expressions of AHR, DAZL, VASA, and PRDM1 in differentiated hESCs treated with 0.1μM DMBA-DHD carrying either control silencing vector shLacZ or shAHR25. Asterisk indicates average from three independent samples, significantly different from respective controls by one-way ANOVA analysis; p < 0.05.

Kehkooi Kee, et al. Toxicol Sci. 2010 September;117(1):218-224.
3.
FIG. 5.

FIG. 5. From: Human Primordial Germ Cell Formation Is Diminished by Exposure to Environmental Toxicants Acting through the AHR Signaling Pathway.

(a) Silencing of AHR specifically rescues VASA:GFP cells. FACS plots of differentiated hESCs carrying VASA:GFP reporter. Percentages of GFP-positive cells are indicated on each plot of 20,000 cells, with the number of VASA:GFP+ in parenthesis. Negative control is hESCs without VASA:GFP reporter. shLacZ or shAHR25 was transduced into hESCs with or without 1μM DMBA-DHD. (b) Histogram plot of the VASA:GFP–positive populations from (a) comparing intensities and number of cells in 20,000 sorted cells.

Kehkooi Kee, et al. Toxicol Sci. 2010 September;117(1):218-224.
4.
FIG. 2.

FIG. 2. From: Human Primordial Germ Cell Formation Is Diminished by Exposure to Environmental Toxicants Acting through the AHR Signaling Pathway.

AHR is silenced in 293FT cells and hESCs. (A) Location of shRNA targeting sequences on the messenger RNA transcript of AHR. (B) Normalized AHR expressions in 293FT cells with control silencing vector, shLacZ, and five shAHR targeting different regions of AHR exons. Two amounts of shAHR, 0.5 and 1 μg, were transfected into 293FT, and qPCR of AHR gene expressions were measured after 24 h. One-microgram shLacZ transfection was used as control and for normalization. Asterisk indicates averages from three independent samples, significantly different from respective controls by one-way ANOVA analysis; p < 0.05.

Kehkooi Kee, et al. Toxicol Sci. 2010 September;117(1):218-224.
5.
FIG. 6.

FIG. 6. From: Human Primordial Germ Cell Formation Is Diminished by Exposure to Environmental Toxicants Acting through the AHR Signaling Pathway.

Caspase 3/7 activity in germ cells is specifically reduced by silencing of AHR. Apoptosis assay measuring Caspase 3/7 activity in FACS cells. Caspase 3/7 activity is triplicate readings of luminescence unit of 1000 sorted cells. Error bars are standard deviations from three measurements. Control is hESCs treated with DMSO and differentiated with BMPs. hESCs carrying shLacZ, shAHR16, or shAHR25 were treated with 1μM DMBA-DHD and differentiated the same way as control. Note that only VASA:GFP+ cells of shLaz had significantly higher Caspase 3/7 activity to all other samples. Asterisk indicates average from three independent samples are significantly different from the respective samples by one-way ANOVA analysis; p < 0.05.

Kehkooi Kee, et al. Toxicol Sci. 2010 September;117(1):218-224.
6.
FIG. 1.

FIG. 1. From: Human Primordial Germ Cell Formation Is Diminished by Exposure to Environmental Toxicants Acting through the AHR Signaling Pathway.

Expression of early germ cell genes is reduced in the presence of PAHs. Normalized gene expression of VASA, DAZL, PRDM1, BAX, NES, and KDR in differentiated hESCs induced by BMPs to germ cells. Controls are differentiated cells treated only with the solvent, DMSO. DMBA, DMBA-DHD, and ANF are dissolved in DMSO before addition to the differentiating cells. Gene expression is measured by qPCR and normalized first to four housekeeping genes (GAPDH, CTNNB1, ACTB, and CENTRIN) followed by normalization to the controls. Error bars are standard deviations from triplicates. Asterisk indicates averages from three independent samples, significantly different from respective controls, by one-way ANOVA analysis; p < 0.05.

Kehkooi Kee, et al. Toxicol Sci. 2010 September;117(1):218-224.

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