Journal: International Journal of Molecular Sciences

Article Title: Glyphosate Exposure Induces Cytotoxicity, Mitochondrial Dysfunction and Activation of ERα and ERβ Estrogen Receptors in Human Prostate PNT1A Cells

doi: 10.3390/ijms25137039

Figure Lengend Snippet: Graphs showing the results of immunofluorescence intensity analysis of ERs in the untreated PNT1A (CTRL) and in cells exposed to Gly (3.5 × 10 −4 M) alone and in the presence of the inhibitor tamoxifen for different times. ( A ) ERα levels. ( B ) ERβ levels. Legend: C—cytoplasm; N—nucleus. Asterisks indicate statistically significant differences between receptor levels in the cytoplasm and nucleus for a specific exposure time: ** p < 0.01; *** p < 0.001, **** p < 0.0001.

Article Snippet: Afterwards, the chambers were incubated for 1 h at room temperature with the primary anti-ERα antibody (E-AB-66893 Elabscience rabbit polyclonal) and the anti-ERβ (SC-8974 Santacruz rabbit polyclonal), both diluted 1:100 in 1% BSA.

Techniques: Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Glyphosate Exposure Induces Cytotoxicity, Mitochondrial Dysfunction and Activation of ERα and ERβ Estrogen Receptors in Human Prostate PNT1A Cells

doi: 10.3390/ijms25137039

Figure Lengend Snippet: Western blot analysis of ERs levels in cytosolic and nuclear protein extracts of PNT1A cells. Representative Western blots of nuclear (N) and cytosolic (C) homogenates from control cells (CTRL) and cells treated for 30′, 2 h, and 4 h with 3.5 × 10 −4 M (LD) Gly, stained with anti-ERα, anti-ERβ, anti-GAPDH, and anti-PARP. ( A , B ) Graphs showing the quantitative results for ERα ( A ) and ERβ ( B ). Nuclear protein level was normalized to PARP levels; cytosolic protein level was normalized to GAPDH levels. Values are means ± SEM of three independent experiments. Asterisks indicate statistically significant differences compared to control cells: ** p < 0.01; *** p < 0.001, **** p < 0.0001.

Article Snippet: Afterwards, the chambers were incubated for 1 h at room temperature with the primary anti-ERα antibody (E-AB-66893 Elabscience rabbit polyclonal) and the anti-ERβ (SC-8974 Santacruz rabbit polyclonal), both diluted 1:100 in 1% BSA.

Techniques: Western Blot, Control, Staining

Journal: International Journal of Molecular Sciences

Article Title: Protecting the Brain: Novel Strategies for Preventing Breast Cancer Brain Metastases through Selective Estrogen Receptor β Agonists and In Vitro Blood–Brain Barrier Models

doi: 10.3390/ijms25063379

Figure Lengend Snippet: Western blot analysis showing the protein expression patterns of the estrogen receptors (ERs) ERα (66 kDa) and ERβ (48 kDa). ( a ) The murine brain endothelial cell lines cEND ( left ) and the human brain endothelial cell line hCMEC/D3 ( right ) show the presence of both ERs in three experimental runs (1, 2, 3). ( b ) The breast cancer (BC) cell lines showed the presence of ERβ in MCF-7, BT-474 and MDA-MB-231 cells, whereas ERα was only detected in MCF-7 and BT-474. Again, three experimental runs were performed (1, 2, 3). β-actin (42 kDa) was used as a loading control.

Article Snippet: We employed the following primary antibodies: ERα (66 kDa, 1:1000, MAB 57151, R&D Systems, Minneapolis, MN, USA), ERβ (48 kDa, 1:250, MAB7106, R&D Systems, Minneapolis, MN, USA), and Cld-5 (18 kDa, 1:500, 35–2500, Invitrogen Thermo Fisher).

Techniques: Western Blot, Expressing, Control