Journal: bioRxiv

Article Title: Discovery of a Selective Inhibitor of ZIP14 with Therapeutic Potential for Cancer-associated Cachexia

doi: 10.1101/2025.10.23.682519

Figure Lengend Snippet: Xenopus oocytes injected either with ( A, B ) human ZIP14 or ( E, F ) human ZIP8 cRNA were incubated with either ( A, E ) ⁶⁵Zn or ( B, F ) ⁵⁵Fe in the presence of ( A, E ) 50 µM or ( B, F ) 10 µM PPTD for 30 min at 22 °C. Radioactivity was measured as described under Methods . Data are normalized against DMSO-treated transporter-expressing controls, and presented as mean ± standard deviation (S.D.) (n=3 independent experiments). Similarly, (C, D) TREx-hZIP14 or (G, H) TREx-hZIP8 cells were treated with Tet (1 μg/mL) for 24 h, followed by incubation either with ( C, G ) ⁵⁴MnCl₂ or (D, H) ¹⁰⁹CdCl₂ in the presence of PPTD at the indicated concentrations for 1 h. Radioactivity was measured as described under Methods . The data (mean + SD) represent results from three independent experiments. Statistical significance was determined using Student’s t -test (A, B, E, F) or one-way ANOVA followed by Dunnett’s post hoc test (C, D, G, H). (* p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001, ***** p < 0.0005).

Article Snippet: The following primary antibodies were used: anti-human ZIP14 (generated previously ), anti-human ZIP8 (Alomone Labs, #AZT-008), and anti-β-actin (Cell Signaling Technology, #3700S).

Techniques: Injection, Incubation, Radioactivity, Expressing, Standard Deviation

Journal: Clinical and Experimental Medicine

Article Title: Unveiling the prognostic and therapeutic landscape of the zinc transporter protein SLC39A family in colorectal cancer through multi-omics and machine learning approaches

doi: 10.1007/s10238-025-01996-2

Figure Lengend Snippet: SLC39A8 and SLC39A14 identified as key model genes playing important roles in tumor microenvironment and cancer progression in CRC (A-B) Representative images of SLC39A8 (A) and SLC39A14 (B) IHC staining in CRC tumors and their paired normal tissues. (C-D) GO and KEGG terms enriched for DEGs between SLC39A8 (C) high/low expressing groups and SLC39A14 (D) high/low expressing groups were identified through GSEA analysis. (E-F) Differences in MsigDB-based pathway activities between SLC39A8 (E) high/low expressing groups and SLC39A14 (F) scored by GSVA. Statistical significance was calculated using Wilcoxon test (E-F). *** p < 0.001, ** p < 0.01, * p < 0.05

Article Snippet: Primary antibodies for SLC39A8 (Cell Signaling Technology, 47188S) and SLC39A14 (Cell Signaling Technology, 24161T) were applied overnight at 4 °C in a humidified chamber.

Techniques: Immunohistochemistry, Expressing

Journal: Annals of Translational Medicine

Article Title: MicroRNA-25-3p therapy for intervertebral disc degeneration by targeting the IL-1β/ZIP8/MTF1 signaling pathway with a novel thermo-responsive vector

doi: 10.21037/atm-20-6595

Figure Lengend Snippet: Different expression profiles of IL-1β, ZIP8, MTF1 and miRNA-25-3p between normal and degenerated nucleus pulposus (NP) tissue. (A) Representative hematoxylin-eosin (HE) and safranin-O-fast green (SO-FG) staining of normal and degenerated human NP tissue. The bar is 500 µm. (B) Relative expression levels of IL-1β mRNA, ZIP8 mRNA and MTF1 mRNA were assessed in normal samples (NS) and degenerated samples (DS) of NP tissue (n=10) via qRT-PCR. (C) Relative expression profiles of IL-1β, ZIP8 and MTF1 proteins were examined in normal and degenerated NP tissue (n=10) via western blotting. (D) Expression of miRNA-25-3p in normal and degenerated NP tissue (n=10). Symbols represent individual disc samples; bars show the mean and 95% confidence interval for each group. *, P<0.05.

Article Snippet: Membranes were incubated overnight at 4 °C with anti-IL-1β antibody (12703, 1:1,000, CST, USA), anti-MTF1 antibody (ab184119, 1:1,000, Abcam, Cambridge, UK), anti-ZIP8 antibody (NBP1-76505, 1:2,000, Novus, Biologicals, Colorado, USA), anti-MMP3 antibody (NB100-91878, 1:1,000, Novus Biologicals, Colorado, USA), anti-MMP13 antibody (NBP2-45887, 1:1,000, Novus, Biologicals, Colorado, USA), anti-ADAMTS5 antibody (NBP2-15286, 1:1,000, Novus, Biologicals, Colorado,USA), anti-β-actin antibody (abs118937, 1:3,000, Absin, Shanghai, China), and anti-PCNA antibody (abs13075, 1:2,000, Absin, Shanghai, China).

Techniques: Expressing, Staining, Quantitative RT-PCR, Western Blot

Journal: Annals of Translational Medicine

Article Title: MicroRNA-25-3p therapy for intervertebral disc degeneration by targeting the IL-1β/ZIP8/MTF1 signaling pathway with a novel thermo-responsive vector

doi: 10.21037/atm-20-6595

Figure Lengend Snippet: Dose-dependent changes in ZIP8 and MTF1 protein expression after IL-1β stimulation in nucleus pulposus (NP) cells. Human NP cells were cultured for 72 hours in serum with different concentrations of IL-1β. (A) ZIP8 protein expression changes with β-actin normalization. (B) Nuclear MTF1 protein expression changes with PCNA normalization. Data are the mean ± standard deviation (SD) of triplicate results. **, P<0.01.

Article Snippet: Membranes were incubated overnight at 4 °C with anti-IL-1β antibody (12703, 1:1,000, CST, USA), anti-MTF1 antibody (ab184119, 1:1,000, Abcam, Cambridge, UK), anti-ZIP8 antibody (NBP1-76505, 1:2,000, Novus, Biologicals, Colorado, USA), anti-MMP3 antibody (NB100-91878, 1:1,000, Novus Biologicals, Colorado, USA), anti-MMP13 antibody (NBP2-45887, 1:1,000, Novus, Biologicals, Colorado, USA), anti-ADAMTS5 antibody (NBP2-15286, 1:1,000, Novus, Biologicals, Colorado,USA), anti-β-actin antibody (abs118937, 1:3,000, Absin, Shanghai, China), and anti-PCNA antibody (abs13075, 1:2,000, Absin, Shanghai, China).

Techniques: Expressing, Cell Culture, Standard Deviation

Journal: Annals of Translational Medicine

Article Title: MicroRNA-25-3p therapy for intervertebral disc degeneration by targeting the IL-1β/ZIP8/MTF1 signaling pathway with a novel thermo-responsive vector

doi: 10.21037/atm-20-6595

Figure Lengend Snippet: MTF1 is a target gene of miRNA-25-3p. (A) Schematic representation of the MTF 3'UTR showing the putative miRNA target site. (B) Luciferase activity of the MTF1 3'UTR reporter was analyzed in nucleus pulposus (NP) cells. An miRNA-25-3p mimic and its negative control (NC) were co-transfected with the wild-type MTF1 3'UTR or mutant vector. (C) The relative MTF1 mRNA and (D) protein expression of transfected NP cells with the miRNA-25-3p mimic and the NC were assessed using qRT-PCR and western blotting, respectively. β-actin was used as an internal control. (E) Illustration depicting the IL-1β/ZIP8/MTF1 signaling pathway regulatory mechanisms and miRNA-25-3p targets. Data are represented as the mean ± standard deviation (SD). *, P<0.05.

Article Snippet: Membranes were incubated overnight at 4 °C with anti-IL-1β antibody (12703, 1:1,000, CST, USA), anti-MTF1 antibody (ab184119, 1:1,000, Abcam, Cambridge, UK), anti-ZIP8 antibody (NBP1-76505, 1:2,000, Novus, Biologicals, Colorado, USA), anti-MMP3 antibody (NB100-91878, 1:1,000, Novus Biologicals, Colorado, USA), anti-MMP13 antibody (NBP2-45887, 1:1,000, Novus, Biologicals, Colorado, USA), anti-ADAMTS5 antibody (NBP2-15286, 1:1,000, Novus, Biologicals, Colorado,USA), anti-β-actin antibody (abs118937, 1:3,000, Absin, Shanghai, China), and anti-PCNA antibody (abs13075, 1:2,000, Absin, Shanghai, China).

Techniques: Luciferase, Activity Assay, Negative Control, Transfection, Mutagenesis, Plasmid Preparation, Expressing, Quantitative RT-PCR, Western Blot, Control, Standard Deviation

Journal: Sleep

Article Title: A potential role for zinc in restless legs syndrome

doi: 10.1093/sleep/zsaa236

Figure Lengend Snippet: Quantification of zinc transporter protein levels in RLS and Control peripheral lymphocytes. The levels of zinc transporter (ZIP8, ZIP14, and TRPM7) were determined with ELISA assays. No significant difference was observed in the protein levels of three zinc transporters. (A) ZIP8 (RLS: n=17, Control: 18); (B) ZIP14 (RLS: n = 15, Control: 17); (C) TPRM7 (RLS: n = 15, Control: 17). The box edges denote the upper and lower quartile range, with the box itself denoting the interquartile range, whiskers denote the full data range, and the horizontal line denotes the median data value for each group. The data were analyzed by Student’s t-test. For each experiment, n > 15 samples measured, and each sample was run in duplicate.

Article Snippet: Quantification of zinc transporter (ZIP14, ZIP8, and TRPM7) was performed using ELISA assays against human ZIP8 (MBS9317558), ZIP14 (MBS9319878), and TRPM7 (MBS9322665) from MyBiosource (San Diego, CA).

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: Blood Advances

Article Title: A mouse model characterizes the roles of ZIP8 in systemic iron recycling and lung inflammation and infection

doi: 10.1182/bloodadvances.2022007867

Figure Lengend Snippet: ZIP8 KO mice have decreased liver manganese and increased splenic iron levels. ICP-MS was performed on liver and spleens from 16-week-old ZIP8 KO mice and littermate controls (fl/fl). n = 5 mice per group. (A,C) Concentrations and (B,D) total levels of (A-B) liver and (C-D) splenic iron, manganese, and zinc were measured. Graphs depict mean and SD; P values determined by 2-tailed unpaired Student t test. SD, standard deviation.

Article Snippet: Monoclonal mouse anti-ZIP8 antibody (9D4A9) was generated and purified by Genscript using the antigen protein sequence: MHPSEGPELAFSEDVLSVFGANRSLSAAQLGRLLERLGAASQQGALDLGQLHFNQCLSAEDIFSLHGFSNVTQITSSNFSAICPAILQQLNFHPCEDLRKHNAKPSHHHHHH.

Techniques: Standard Deviation

Journal: Blood Advances

Article Title: A mouse model characterizes the roles of ZIP8 in systemic iron recycling and lung inflammation and infection

doi: 10.1182/bloodadvances.2022007867

Figure Lengend Snippet: Baseline ZIP8 KO mice have impaired iron recycling. (A-D) Tissues and serum from 16-week-old ZIP8 KO and littermate controls (fl/fl) were harvested for analysis. n = 4 to 14 mice per group. (A) Liver, spleen and serum nonheme iron levels. (B) CBCs. (C) Liver Hamp mRNA and BM Erfe mRNA levels. (D) Representative images of Perls’ staining for iron in ZIP8 KO and fl/fl FFPE spleen tissue. 40× original magnification. (E) Splenic red pulp macrophages from C57BL/6 mice were isolated via magnetic bead selection for F4/80+ cells. ZIP8 protein levels are enriched in the splenic F4/80+ population as determined by western blot. n = 4 mice per group. (A-C) Graphs depict mean and SD; P values determined by 2-tailed unpaired Student t test. BM, bone marrow; FFPE, formalin-fixed paraffin-embedded; SD, standard deviation.

Article Snippet: Monoclonal mouse anti-ZIP8 antibody (9D4A9) was generated and purified by Genscript using the antigen protein sequence: MHPSEGPELAFSEDVLSVFGANRSLSAAQLGRLLERLGAASQQGALDLGQLHFNQCLSAEDIFSLHGFSNVTQITSSNFSAICPAILQQLNFHPCEDLRKHNAKPSHHHHHH.

Techniques: Staining, Isolation, Selection, Western Blot, Formalin-fixed Paraffin-Embedded, Standard Deviation

Journal: Blood Advances

Article Title: A mouse model characterizes the roles of ZIP8 in systemic iron recycling and lung inflammation and infection

doi: 10.1182/bloodadvances.2022007867

Figure Lengend Snippet: ZIP8 KO and WT mice have similar responses to PHZ. (A-C) C57BL/6 mice were treated with 300 μL of aged RBCs, a single IP injection of 10 mg iron dextran or 2 consecutive daily doses of 60 mg/kg PHZ to induce hemolytic anemia. n = 3 to 5 mice for each treatment group. Tissues were harvested 3 days after treatment. n = 3 to 5 mice per group. (A) Western blot of splenic ZIP8. (B) Correlation between splenic ZIP8 protein levels as quantified by densitometry and splenic nonheme iron levels. P value and R 2 as determined by Pearson’s correlation. (C) Liver Saa1 mRNA levels. (D-F) ZIP8 KO and littermate controls (fl/fl) were treated with PHZ as described above. n = 6 to 9 mice per group. (D) CBCs were performed on day 4 and day 7 on heparinized blood from ZIP8 KO and fl/fl mice. (E) Day 7 liver Saa1 , liver Nqo1 , and spleen Nqo1 mRNA levels and (F) spleen and serum nonheme iron. (A,C-F) Graphs depict mean and SD; P values determined by 2-tailed unpaired Student t test. SD, standard deviation; WT, wild-type.

Article Snippet: Monoclonal mouse anti-ZIP8 antibody (9D4A9) was generated and purified by Genscript using the antigen protein sequence: MHPSEGPELAFSEDVLSVFGANRSLSAAQLGRLLERLGAASQQGALDLGQLHFNQCLSAEDIFSLHGFSNVTQITSSNFSAICPAILQQLNFHPCEDLRKHNAKPSHHHHHH.

Techniques: Injection, Western Blot, Standard Deviation

Journal: Blood Advances

Article Title: A mouse model characterizes the roles of ZIP8 in systemic iron recycling and lung inflammation and infection

doi: 10.1182/bloodadvances.2022007867

Figure Lengend Snippet: Loss of ZIP8 has no effect on response to iron deficiency in mice. ZIP8 KO mice and littermate controls (fl/fl) were placed on a 4 ppm Fe diet and CBCs were measured at 6, 8, 12, 16, 20, and 24 weeks on the diet. Tissues and serum were harvested at the end of the treatment. n = 8 to 9 mice per group. (A) CBCs from ZIP8 KO and fl/fl mice. (B) Blood ZPP levels. (C) Spleen, liver, and serum nonheme iron levels from iron-deficient ZIP8 KO mice and fl/fl mice. (D) Liver Hamp1 mRNA. (E) Representative western blot for splenic Fpn, Tfr1, and FtnH protein. (A-D) Graphs depict mean and SD. SD, standard deviation; ZPP, zinc protoporphyrin.

Article Snippet: Monoclonal mouse anti-ZIP8 antibody (9D4A9) was generated and purified by Genscript using the antigen protein sequence: MHPSEGPELAFSEDVLSVFGANRSLSAAQLGRLLERLGAASQQGALDLGQLHFNQCLSAEDIFSLHGFSNVTQITSSNFSAICPAILQQLNFHPCEDLRKHNAKPSHHHHHH.

Techniques: Western Blot, Standard Deviation

Journal: Blood Advances

Article Title: A mouse model characterizes the roles of ZIP8 in systemic iron recycling and lung inflammation and infection

doi: 10.1182/bloodadvances.2022007867

Figure Lengend Snippet: ZIP8 is induced by inflammation in human alveolar epithelial cells. (A) Representative images for IHC for ZIP8 (brown) and SP-C (red; AT2 cells) in normal human lung tissue (n = 2 biological replicates). Images are 10× (left) and 40× (right) original magnification. (B-D) A549 cells (human alveolar epithelial cell line) were serum starved for 24 hours, then treated with 50 ng/mL recombinant human IL-6, IFN-γ, IL-1α, IL-1β, TNFα, or epidermal growth factor for 16 hours. n = 1 to 3 independent experiments. (B) Representative western blot for ZIP8 protein expression. (C) SLC39A8 mRNA levels. (D) ZIP8 siRNA knockdown. (E) 16HBE cells (human bronchiolar epithelial cell line) SLC39A8 mRNA levels after treatment for 16 hours. Graphs depict mean and SD. IFN- γ, interferon gamma; KD, ZIP8 siRNA; NT, nontargeting siRNA; SD, standard deviation.

Article Snippet: Monoclonal mouse anti-ZIP8 antibody (9D4A9) was generated and purified by Genscript using the antigen protein sequence: MHPSEGPELAFSEDVLSVFGANRSLSAAQLGRLLERLGAASQQGALDLGQLHFNQCLSAEDIFSLHGFSNVTQITSSNFSAICPAILQQLNFHPCEDLRKHNAKPSHHHHHH.

Techniques: Recombinant, Western Blot, Expressing, Knockdown, Standard Deviation

Journal: Blood Advances

Article Title: A mouse model characterizes the roles of ZIP8 in systemic iron recycling and lung inflammation and infection

doi: 10.1182/bloodadvances.2022007867

Figure Lengend Snippet: ZIP8 transports iron from the airspace into the lung. (A) Slc39a8 mRNA levels of C57BL/6 mouse lung subpopulation cell types. B CD45+: BAL macrophages. L CD45+: lung macrophages. CD31+: endothelial cells. Epcam+: epithelial cells. n = 3 to 4 mice per group. (B) ZIP8 KO mice and littermate controls (fl/fl) were treated with 5 μg of 58 Fe 2+ via RO injection or OP aspiration and harvested after 4 hours for ICP-MS analysis of 58 Fe levels. n = 3 mice per group. (C-D) 16-week-old ZIP8 KO and fl/fl mouse lung (C) nonheme iron levels and (D) Nqo1 mRNA and MDA levels. n = 4 to 10 mice per group. (A-D) Graphs depict mean and SD; P values were determined by 2-tailed unpaired Student t test. MDA, malondialdehyde; SD, standard deviation.

Article Snippet: Monoclonal mouse anti-ZIP8 antibody (9D4A9) was generated and purified by Genscript using the antigen protein sequence: MHPSEGPELAFSEDVLSVFGANRSLSAAQLGRLLERLGAASQQGALDLGQLHFNQCLSAEDIFSLHGFSNVTQITSSNFSAICPAILQQLNFHPCEDLRKHNAKPSHHHHHH.

Techniques: Injection, Standard Deviation

Journal: Blood Advances

Article Title: A mouse model characterizes the roles of ZIP8 in systemic iron recycling and lung inflammation and infection

doi: 10.1182/bloodadvances.2022007867

Figure Lengend Snippet: ZIP8 does not play a significant role in LPS-induced ALI. 16-week-old ZIP8 KO mice and littermate controls (fl/fl) were treated with 15 mg/kg LPS through OP aspiration to induce ALI and harvested after 3 days. A subset of mice in both groups were left untreated for comparison. n = 3 to 7 mice per group. (A) Representative western blot for ZIP8 in lung lysate. (B) Lung nonheme iron levels. (C) BAL protein levels. (D) BAL and lung tissue MPO activity. (E) Lung mitochondrial SOD. (B-E) Graphs depict mean and SD. SD, standard deviation.

Article Snippet: Monoclonal mouse anti-ZIP8 antibody (9D4A9) was generated and purified by Genscript using the antigen protein sequence: MHPSEGPELAFSEDVLSVFGANRSLSAAQLGRLLERLGAASQQGALDLGQLHFNQCLSAEDIFSLHGFSNVTQITSSNFSAICPAILQQLNFHPCEDLRKHNAKPSHHHHHH.

Techniques: Comparison, Western Blot, Activity Assay, Standard Deviation

Journal: Blood Advances

Article Title: A mouse model characterizes the roles of ZIP8 in systemic iron recycling and lung inflammation and infection

doi: 10.1182/bloodadvances.2022007867

Figure Lengend Snippet: ZIP8 deletion is protective against K pneumoniae infection. ZIP8 KO mice and littermate controls (fl/fl) were infected with 1500 CFU of K pneumoniae via OP aspiration and weighed daily until harvest at 3 days. (A) Percent weight loss in ZIP8 KO and fl/fl mice. n = 3 to 4 mice per group. (B) CFU counts in lung, liver, spleen, and blood after infection. (C) BAL protein levels. (D) BAL and lung tissue MPO activity. (E) Lung mitochondrial and cytosolic SOD. Graphs depict mean and SD; P values were determined by 2-tailed unpaired Student t test. SD, standard deviation.

Article Snippet: Monoclonal mouse anti-ZIP8 antibody (9D4A9) was generated and purified by Genscript using the antigen protein sequence: MHPSEGPELAFSEDVLSVFGANRSLSAAQLGRLLERLGAASQQGALDLGQLHFNQCLSAEDIFSLHGFSNVTQITSSNFSAICPAILQQLNFHPCEDLRKHNAKPSHHHHHH.

Techniques: Infection, Activity Assay, Standard Deviation