Ferroptosis Mechanism of HepG2 Cells

HepG2 cell line was obtained from HonorGene (Changsha, China). Ferrostatin-1, MTT, 3-methyladenin (3-MA), chloroquine (CQ), RPMI-1640, and other chemicals were purchased from Sigma-Aldrich. 2-Pyridylhydrazone dithiocarbamate s-acetic acid (PdtaA) was prepared in our laboratory [27 (link)]. GPx4, xCT (SLC7A11), NDRG1, vimentin, and NCOA4 antibody were obtained from the Proteintech Group (Wuhan, China). Antibodies of E-cadherin and secondary antibodies (fluorescence-labeled for immunofluorescence) were purchased from Cell Signaling Technology (USA). Ferritin antibody for immunofluorescence was obtained from Santa Cruz Biotechnology (USA, Santa Cruz). NCOA4 antibody for immunofluorescence was purchased from Atlas Antibody (Sweden). Secondary antibodies for Western blotting were obtained from EarthOx, LLC (San Francisco, USA).

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Li H., Zhou W., Wei H., Li L., Wang X., Li Y., Li S, & Li C. (2021). Ferritinophagic Flux Was a Driving Force in Determination of Status of EMT, Ferroptosis, and NDRG1 Activation in Action of Mechanism of 2-Pyridylhydrazone Dithiocarbamate S-Acetic Acid. Journal of Oncology, 2021, 3015710.

Publication 2021

Ferrostatin-1 MTT 3-methyladenin (3-MA), chloroquine (CQ), RPMI-1640 vimentin NCOA4 Ferritin antibody for immunofluorescence Secondary antibodies for Western blotting

Acetic acid Antibodies Antibody Cadherin Chloroquine Ferritin Ferrostatin 1 Fluorescence Gpx4 Hepg2 cell line Immunofluorescence Ndrg1 Vimentin

Corresponding Organization : Xinxiang Medical University

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Variable analysis

independent variables

Ferrostatin-1
3-methyladenin (3-MA)
Chloroquine (CQ)
2-Pyridylhydrazone dithiocarbamate s-acetic acid (PdtaA)

dependent variables

Cell viability (measured by MTT assay)
Levels of GPx4, xCT (SLC7A11), NDRG1, vimentin, NCOA4, E-cadherin, and Ferritin (measured by Western blotting and/or immunofluorescence)

control variables

RPMI-1640 cell culture medium
HepG2 cell line

controls

Positive control: None specified
Negative control: None specified

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