Journal: Journal of the Endocrine Society

Article Title: Estrogenic activity in tampon products

doi: 10.1210/jendso/bvag094

Figure Lengend Snippet: Estradiol dose-response and recovery of spiked estradiol from tampon extracts. (A) T47D-luc cells were treated with an 11-point estradiol dilution series starting at 30nM, followed by 10nM, 3.3nM, 1.11nM, 370pM, 123pM, 41pM, 14pM, 4.6pM, 1.5pM, and 0.5pM). For each concentration, 2 μL of steroid dilution was added to 198 μL assay medium, and cells were incubated for 24 hours. Estrogen-responsive luciferase activity was quantified using BrightGlo substrate. Luminescence values represent the mean of 4 technical replicates, and a sigmoidal dose-response curve with EC 50 determination was generated using GraphPad v10. (B) Recovery of estradiol from tampon brands A-D following spiking and extraction. One gram of each tampon from Brands A, B, C, and D were spiked with 2nM estradiol, soaked in a 1:1 water:ethanol solution for 4 hours, extracted, evaporated to dryness, and reconstituted in methanol. Extracts were diluted 1:100 in cell culture medium and then serially diluted 1:2 to generate 4 assay points. T47D-luc cells were incubated with a further 1:10 dilution of each tampon extract for 24 hours and luciferase activity measured using BrightGlo. Each point represents mean ± SEM of 4 technical replicates corrected for the blank control sample (1:1 ethanol:water solution SPE-extracted as per samples). The data for the spiked tampons are plotted alongside estradiol standards run in parallel.

Article Snippet: Estrogen Receptor Luciferase Reporter T47D Stable Cell Line cells (Signosis, Santa Clara, CA, USA: Cat # SL-0002, T47D-luc)) were grown to 90% confluence in a 550 mL cell culture flask with RPMI 164 medium supplemented with 10% (v/v) Hyclone fetal bovine serum (GE Lifesciences) and Geneticin (75 μg mL −1 , G148, Life Technologies).

Techniques: Concentration Assay, Incubation, Luciferase, Activity Assay, Generated, Extraction, Cell Culture, Control

Journal: Journal of the Endocrine Society

Article Title: Estrogenic activity in tampon products

doi: 10.1210/jendso/bvag094

Figure Lengend Snippet: Estrogenic activity detected in extracts from 8 commercially available tampon brands (A-H). T47D-luc cells were exposed to methanolic extracts prepared from 1 g of core tampon material (string removed) from 8 tampon brands sold in New Zealand. Each 1 g sample was soaked in a 1:1 ethanol:water solution for 4 hours, sonicated, filtered, extracted, and then evaporated to dryness, and reconstituted in methanol. The final extract was diluted into assay medium at 1:1000 (1), followed by serial dilutions to generate 1:2000 (2), 1:4000 (3), and 1:8000 (4) exposure concentrations. Cells were incubated with each dilution for 24 hours, and estrogen-responsive luciferase activity was quantified using a luminescence plate reader. Data are presented as the mean ± SEM of 2 independently extracted tampons per brand, each tested as 2 portions of 1 g each, and each dilution tested with 4 technical replicates.

Article Snippet: Estrogen Receptor Luciferase Reporter T47D Stable Cell Line cells (Signosis, Santa Clara, CA, USA: Cat # SL-0002, T47D-luc)) were grown to 90% confluence in a 550 mL cell culture flask with RPMI 164 medium supplemented with 10% (v/v) Hyclone fetal bovine serum (GE Lifesciences) and Geneticin (75 μg mL −1 , G148, Life Technologies).

Techniques: Activity Assay, Sonication, Incubation, Luciferase, Microplate Reader Luminescence Measurement

Journal: Journal of the Endocrine Society

Article Title: Estrogenic activity in tampon products

doi: 10.1210/jendso/bvag094

Figure Lengend Snippet: Independent batches of Tampon Brand D show estrogenic activity. T47D-luc cells were treated with methanolic extracts generated from 1 g of core tampon material (string removed) from 3 independent batches of Brand A and Brand D. For each batch, 3 separate tampons were extracted independently. Each 1 g sample was soaked in 1:1 ethanol:water solution for 4 hours, sonicated, filtered, extracted, and evaporated to dryness, and reconstituted in methanol. Reconstituted extracts were diluted into assay medium at 1:1000 (1), followed by serial 1:2 dilutions to produce 1:2000 (2), 1:4000 (3), and 1:8000 (4) exposure concentrations. Cells were incubated for 24 hours, and estrogen-responsive luciferase activity was quantified by using a luminescence plate reader. Data are shown as mean ± SEM of 3 independently extracted tampons per batch, tested as 2 portions of 1 g each with each dilution tested as 4 technical replicates.

Article Snippet: Estrogen Receptor Luciferase Reporter T47D Stable Cell Line cells (Signosis, Santa Clara, CA, USA: Cat # SL-0002, T47D-luc)) were grown to 90% confluence in a 550 mL cell culture flask with RPMI 164 medium supplemented with 10% (v/v) Hyclone fetal bovine serum (GE Lifesciences) and Geneticin (75 μg mL −1 , G148, Life Technologies).

Techniques: Activity Assay, Generated, Sonication, Incubation, Luciferase, Microplate Reader Luminescence Measurement

Journal: Journal of the Endocrine Society

Article Title: Estrogenic activity in tampon products

doi: 10.1210/jendso/bvag094

Figure Lengend Snippet: Estrogenic activity of internationally sourced tampon brands. T47D-luc cells were treated with methanolic extracts prepared from intact tampons (string removed) from a panel of internationally sourced tampon brands (Brands I-R). For each brand, 2 or 3 independent tampons were extracted separately on independent days. Each tampon was soaked in a 1:1 ethanol:water solution for 4 hours, sonicated, filtered, extracted, evaporated to dryness, and reconstituted in methanol. Reconstituted extracts were diluted into assay medium at 1:1000, and cells were incubated for 24 hours before estrogen-responsive luciferase activity was quantified using a luminescence plate reader. Data are shown as mean ± SEM of 2 or 3 independently extracted tampons per brand with each dilution tested as 4 technical replicates.

Article Snippet: Estrogen Receptor Luciferase Reporter T47D Stable Cell Line cells (Signosis, Santa Clara, CA, USA: Cat # SL-0002, T47D-luc)) were grown to 90% confluence in a 550 mL cell culture flask with RPMI 164 medium supplemented with 10% (v/v) Hyclone fetal bovine serum (GE Lifesciences) and Geneticin (75 μg mL −1 , G148, Life Technologies).

Techniques: Activity Assay, Sonication, Incubation, Luciferase, Microplate Reader Luminescence Measurement

Journal: Journal of the Endocrine Society

Article Title: Estrogenic activity in tampon products

doi: 10.1210/jendso/bvag094

Figure Lengend Snippet: Effect of sonication on estrogenic activity of tampon extracts. T47D-luc cells were exposed to methanolic extracts prepared from 2 independent tampons per brand (Brands A and D, string removed), comparing extraction with and without sonication. Each tampon was soaked in a 1:1 ethanol:water solution for 4 hours, either sonicated or left nonsonicated, then filtered, extracted, evaporated to dryness, and reconstituted in methanol. Extracts were diluted into assay medium at 1:1000 (1), followed by serial 1:2 dilutions to generate 1:2000 (2) and 1:4000 (3) exposure concentrations. Cells were incubated for 24 hours, and estrogen-responsive luciferase activity was quantified. Data represent mean ± SEM of t2wo independently extracted tampons per condition with each dilution tested as 4 technical replicates.

Article Snippet: Estrogen Receptor Luciferase Reporter T47D Stable Cell Line cells (Signosis, Santa Clara, CA, USA: Cat # SL-0002, T47D-luc)) were grown to 90% confluence in a 550 mL cell culture flask with RPMI 164 medium supplemented with 10% (v/v) Hyclone fetal bovine serum (GE Lifesciences) and Geneticin (75 μg mL −1 , G148, Life Technologies).

Techniques: Sonication, Activity Assay, Extraction, Incubation, Luciferase

Journal: bioRxiv

Article Title: Mitochondrial inner membrane interactors of Aurora kinase A/AURKA and PHB2 shape organelle metabolic heterogeneity

doi: 10.64898/2026.03.17.712317

Figure Lengend Snippet: Upper panels) Maximal projections of representative 2D dSTORM super-resolution micrographs from T47D cells treated with DMSO or HMBB for 24 h, and co-stained for ATP5F1B and MIC60. The anti-ATP5F1B primary antibody was detected using an Alexa 647-conjugated secondary antibody and pseudocolored cyan, while the anti-MIC60 primary antibody was detected using a CF532 secondary antibody and pseudocolored magenta. Insets: magnified region where each staining is merged (ZOOM). Scale bar: 10 μ m. (Lower panel) Colocalization analyses performed with the GCoPS software, showing the percentage of colocalization between MIC60 and ATP5F1B in controls and HMBB-treated cells. n = 10 cells per condition in a representative experiment (of two). Data are means ± S.D. ns: not significant.

Article Snippet: MCF7 (HTB-22) and T47D (HTB-133) cells were purchased from the American Type Culture Collection, while Flp-InTM T-RExTM HEK293 cells were purchased from Thermo Fisher Scientific.

Techniques: Staining, Software

Journal: bioRxiv

Article Title: Mitochondrial inner membrane interactors of Aurora kinase A/AURKA and PHB2 shape organelle metabolic heterogeneity

doi: 10.64898/2026.03.17.712317

Figure Lengend Snippet: ( A ) Relative mitochondrial area shown as the amount of PMPCB staining (threshold mask and corresponding quantification) in T47D cells transfected as indicated. A PMPCB : mitochondrial area normalized against total cell area (%). n = 10 cells per condition (small dots) in each of three experimental replicates. Large dots indicate mean values for each replicate. Data are means ± S.D. Scale bar: 10 µm. ( B-C ) Representative western blot and corresponding quantifications of the relative abundance of PMPCB, SLC25A13 ( B ), and PPARGC1 ( C ) in total lysates of T47D cells transfected with a control or SLC25A13 -specific siRNAs. Loading controls: TUBA1A ( B ), ACTA ( C ). Data are from n = 3 or 4 independent experiments, and presented as means ± S.D. A.U.: arbitrary units. * P < 0.05, ** P < 0.01. ns: not significant.

Article Snippet: MCF7 (HTB-22) and T47D (HTB-133) cells were purchased from the American Type Culture Collection, while Flp-InTM T-RExTM HEK293 cells were purchased from Thermo Fisher Scientific.

Techniques: Staining, Transfection, Western Blot, Control

Journal: bioRxiv

Article Title: Mitochondrial inner membrane interactors of Aurora kinase A/AURKA and PHB2 shape organelle metabolic heterogeneity

doi: 10.64898/2026.03.17.712317

Figure Lengend Snippet: ( A ) (From left to right) eSRRF-reconstructed images of GFP, mKO2 (FRET), ratiometric FRET images and visual representation of ratiometric FRET shown in greyscale upon random FRET analyses of T47D cells expressing the mitoGO-ATeam2 biosensor and transfected with control or SLC25A13 -specific siRNAs, and treated with DMSO or HMBB as indicated. Lines corresponding to FRET Hotspots or Cordpots are pseudocolored in red and cyan, respectively. Insets: higher magnification of the squared area. Graph: mean FRET index values (0-100%) in the indicated conditions. n = 10 cells per condition (small dots) in each of three experimental replicates. Large dots indicate mean values for each replicate. Data are means ± S.D. Scale bar: 5 µm. ( B ) Table showing the overall repartition of FRET Coldspots and Hotspots detected with the MitoGO-ATeam2 biosensor in the indicated transfection condition, together with the quantity of FRET Coldspots and Hotspots depending on the degree of variation of the FRET index (clusters). For each cluster and spot type, the FRET index peak value is shown in red (FRET Hotspots) or cyan (FRET Coldspots). ( C ) Representative western blot and corresponding quantifications of the relative abundance of PPARGC1 in total lysates of T47D cells treated with DMSO or HMBB for 24 h. Loading control: ACTA. Data are from n = 3 independent experiments, and presented as means ± S.D. A.U.: arbitrary units. * P < 0.05, ** P < 0.01. ns: not significant.

Article Snippet: MCF7 (HTB-22) and T47D (HTB-133) cells were purchased from the American Type Culture Collection, while Flp-InTM T-RExTM HEK293 cells were purchased from Thermo Fisher Scientific.

Techniques: Expressing, Transfection, Control, Western Blot