Characterization of Zip7 Inhibitors

NVS-ZP7-4 is an inhibitor of Zip7 that was derived from a compound (NVS-ZP7-1) that was obtained in a phenotypic screen of inhibitors of Notch signaling as previously described [23 (link)]. NVS-ZP7-6 is an inactive analog of NVS-ZP7-4. NVS-ZP7-1, NVS-ZP7-4 and NVS-ZP7-6 were synthesized internally at Novartis. Tauroursodeoxycholic acid (TUDCA) (cat# T0266), salubrinal (cat# 324895), dibenzazepine (cat# 209984-56-S) and compound E (cat# 209986-17-4) were purchased from Sigma. All recombinant proteins were purchased from R&D Systems, Minneapolis, MN, unless otherwise noticed. TPEN (N,N,N′,N′-Tetrakis(2-pyridylmethyl)ethylenediamine) was purchased from Sigma (cat# P4413). DTPA (diethylenetriaminepentaacetic acid) was purchased from Sigma (cat# D1133). The following antibodies were purchased from Cell Signaling Technologies and used at 1:1000 dilution: BiP (C50B12) rabbit mAb #3177, IRE1 α (14C10) rabbit mAb #3294, CHOP (L63F7) mouse mAb #2985, calreticulin (D3E6) rabbit mAb #12238, PERK (C33E10) rabbit mAb #3192, XBP-1s (D2C1F) rabbit mAb #12782, ZIP7/SLC39A& (D103A) rabbit mAb # 33176. The following antibodies were purchased from Cell Technologies and used at 1:2000 dilution: GAPDH (14C10) rabbit mAb #2118, β -actin (8H10D10) mouse mAb #3700, goat anti-rabbit HRP-linked IgG #7074, horse anti-mouse HRP-linked IgG #7076.

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Xu Y., Twarog M., Li N., Banks A., Schustak J., Bao Y., Huang Q, & Medley Q.G. (2022). Zinc transport from the endoplasmic reticulum to the cytoplasm via Zip7 is necessary for barrier dysfunction mediated by inflammatory signaling in RPE cells. PLoS ONE, 17(7), e0271656.

Publication 2022

Tauroursodeoxycholic acid (TUDCA) salubrinal dibenzazepine compound E

Actin Anti igg Antibodies Calreticulin Cell Chop Compound e Dibenzazepine Diethylenetriaminepentaacetic acid Dilution Ethylenediamine Gapdh Goat Horse Inhibitors Mouse Nvs 1 Phenotypic Rabbit Recombinant proteins Salubrinal Tauroursodeoxycholic acid Tpen

Corresponding Organization : Novartis (United States)

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Variable analysis

independent variables

NVS-ZP7-4 (Zip7 inhibitor)
NVS-ZP7-6 (inactive analog of NVS-ZP7-4)
Tauroursodeoxycholic acid (TUDCA)
Salubrinal
Dibenzazepine
Compound E
TPEN (N,N,N′,N′-Tetrakis(2-pyridylmethyl)ethylenediamine)
DTPA (diethylenetriaminepentaacetic acid)

dependent variables

Notch signaling
Zip7 activity
Expression of ER stress markers (BiP, IRE1α, CHOP, calreticulin, PERK, XBP-1s, ZIP7/SLC39A7)

control variables

Cell culture conditions
Protein purification and characterization methods
Antibody dilutions and sources

controls

Positive control: NVS-ZP7-1 (compound obtained in a phenotypic screen for Notch signaling inhibitors)
Negative control: NVS-ZP7-6 (inactive analog of NVS-ZP7-4)

Annotations

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