rabbit anti fsp1  (Proteintech)

Journal: Nature Communications

Article Title: Suppression of ferroptosis by vitamin A or radical-trapping antioxidants is essential for neuronal development

doi: 10.1038/s41467-024-51996-1

Figure Lengend Snippet: a Flow cytometry of C11-BODIPY staining: (left) histograms and (right) intensity of HT-1080 cells co-treated with the ferroptosis inducer (RSL3), and inhibitor (Fer-1) or vitamin A (ATRA). Data are % intensity of median fluorescence normalized to RSL3-treated cells ± SD of n = 4 biologically independent replicates; one-way-ANOVA with Tukey’s test. b Immunostaining of 4-Hydroxynonenal (4-HNE) and flow cytometry: (left) histograms and (right) intensity of HT-1080 cells co-treated with the ferroptosis inducer (RSL3), and inhibitor (Fer-1) or vitamin A (ATRA). Data are % intensity of median fluorescence normalized to RSL3-treated cells ± SD of n = 3 biologically independent replicates; one-way-ANOVA with Tukey’s test. c TBARS assay of HT-1080 cells co-treated with the ferroptosis inducer (RSL3) and inhibitor (Fer-1) or vitamin A (ATRA). Data are mean ± SD of n = 4 biologically independent replicates; one-way-ANOVA with Tukey’s test. d mRNA expression, using quantitative RT-PCR, of various anti-ferroptotic regulators in HT-1080 cells treated with DMSO or 10 µM vitamin A (ATRA) and 10 µM RARi. Data are mean ± SD of n = 3 biologically independent replicates; one-way-ANOVA with Tukey’s test. e protein expression, using Western Blot, of various anti-ferroptotic regulators (GPX4, FSP1, ACSL3, SCD1) as well as pro-ferroptotic regulators (ACSL4, LPCAT3) in HT-1080 cells treated with DMSO or 20 µM vitamin A (ATRA). n = 3 biologically independent replicates are shown. f mRNA expression (qRT-PCR) of various anti-ferroptotic regulators in cortical neurons differentiated with low or high vitamin A. Data plotted are mean ± SD of n = 3 biologically independent replicates; unpaired t test, two-tailed. g protein expression, using Western Blot, of various anti-ferroptotic regulators (GPX4, FSP1, SCD1) in immature neurons differentiated with low or high vitamin A. n = 3 biologically independent replicates are shown. Source data are provided as a Source Data file.

Article Snippet: The membrane was blocked with 5% milk in TBS-Tween and afterward incubated overnight at 4 °C in the following primary antibodies: rabbit anti-GPX4 (ab125066, Abcam, 1:1000), rabbit anti-FSP1 (AMID, PA5-103183, Thermo Fisher Scientific, 1:500), rabbit anti-ACSL3 (ab151959, Abcam, 1:1000), rabbit anti-SCD1 (ab236868, Abcam, 1:1000), mouse anti-ACSL4 (sc-365230, Santa Cruz Biotechnology, 1:500), mouse anti-LPCAT3 (ab239585, Abcam, 1:1000).

Techniques: Flow Cytometry, Staining, Fluorescence, Immunostaining, TBARS Assay, Expressing, Quantitative RT-PCR, Western Blot, Two Tailed Test

Journal: Nature Communications

Article Title: Suppression of ferroptosis by vitamin A or radical-trapping antioxidants is essential for neuronal development

doi: 10.1038/s41467-024-51996-1

Figure Lengend Snippet: This graphical summary shows that lipid peroxidation and ferroptosis must be suppressed to generate mature neurons. This is achieved either through the action of radical-trapping antioxidants (RTAs) or by transcriptional up-regulation of ferroptosis gatekeepers GPX4, FSP1, GCH1, and ACSL3 by all-trans retinoic acid (vitamin A). Vitamin A has a dual function in suppressing ferroptosis as both retinal and retinol have RTA activity.

Article Snippet: The membrane was blocked with 5% milk in TBS-Tween and afterward incubated overnight at 4 °C in the following primary antibodies: rabbit anti-GPX4 (ab125066, Abcam, 1:1000), rabbit anti-FSP1 (AMID, PA5-103183, Thermo Fisher Scientific, 1:500), rabbit anti-ACSL3 (ab151959, Abcam, 1:1000), rabbit anti-SCD1 (ab236868, Abcam, 1:1000), mouse anti-ACSL4 (sc-365230, Santa Cruz Biotechnology, 1:500), mouse anti-LPCAT3 (ab239585, Abcam, 1:1000).

Techniques: Activity Assay

Journal: Frontiers in Toxicology

Article Title: Characterization of a human thyroid microtissue model for testing thyroid disrupting chemicals

doi: 10.3389/ftox.2024.1408808

Figure Lengend Snippet: Characterization of the cell population from five donor lots determined by FLOW. (A) Percentage of cells stained positive for EpCAM, CD90, and CD144. > 0.5 × 10 6 viable cells were used for each of the five donor lots. (B) Representative images of thyrocyte cells from donor lots 2217737 (top row) and 2214495 (bottom row) in 2D culture stained for Cytokeratin 7 (CK7, green), Fibroblast-specific protein 1 (FSP1, red), and DAPI (blue), and three channels merged. Magnification ×50. Scale bar = 200 µm. (C) Representative images of microtissues in 3D culture seeded at 7,500 cells per well and (D) levels of Thyroxine (T 4 ) from microtissues in 3D culture on day 14. Cells were treated with Thyroid Stimulating Hormone (1.0 mIU/mL). Magnification ×40. Scale bar = 50 µm. Error bars represent standard deviation. n ≥ 3 replicates from five donor lots.

Article Snippet: Primary antibodies were added including monoclonal mouse anti-human CK7 (Clone OV-TL, Agilent Dako, Santa Clara, CA) (1:100 dilution) and rabbit polyclonal FSP1 (Abcam, Boston, MA) (1:100 dilution), and incubated overnight at 4°C.

Techniques: Staining, Standard Deviation