nb4  (DSMZ)

Journal: Cell reports

Article Title: ATP7B-maintained copper stores in myeloid progenitors are required for functional maturation of neutrophils

doi: 10.1016/j.celrep.2026.116955

Figure Lengend Snippet: (A) Co-immunostaining of ATP7B (green) with the trans-Golgi marker TGN46 (red) in mouse myelocytes and co-immunostaining of ATP7B (green) with the cis-Golgi marker GM130 (red) in human myeloid NB4 cells. A zoomed view used to demonstrate the ATP7B (green) vesicular pattern. (B) Co-immunostaining of ATP7B (yellow) with TGN46 (cyan), the early endosome marker EEA1 (cyan), the lysosome marker LAMP1 (magenta), and the late endosomal marker Rab8 (magenta) in non-differentiated NB4 cells. There was no significant co-localization of ATP7B with Golgi and endosomal markers, respectively. Scale bar, 5 μm. (C) Live images of CopperGREEN sensor in non-differentiated NB4 cells. DAPI (blue) was used as a nuclear marker. Scale bar, 10 μm. See also – .

Article Snippet: Human myeloid NB4 (Cytion, cat# 300299) and NB4/Luc+ cells (AML-M3, gift from Gabriel Ghiaur’s lab) were cultured in RPMI1640 media supplemented with 10% FBS +2 mM Glutamine +1% Pen-Strep.

Techniques: Immunostaining, Marker

Journal: Cell reports

Article Title: ATP7B-maintained copper stores in myeloid progenitors are required for functional maturation of neutrophils

doi: 10.1016/j.celrep.2026.116955

Figure Lengend Snippet: (A) Schematic representation of ATRA-induced human myeloid NB4 cell differentiation. (B) Western blot analysis reveals that ATP7B protein abundance is significantly reduced and CEBPε is upregulated during ATRA-induced NB4 cell differentiation from day 0 to day 6. β-actin was used as an endogenous loading control. Immunoblotting data represent 4–5 independent experiments. (C) Densitometric analysis demonstrating altered protein abundance of ATP7B and CEBPε was performed using ImageJ by normalizing to β-actin. (D) Western blot analysis reveals that ATP7A protein abundance is upregulated during ATRA-induced NB4 cell differentiation from day 0 to day 6. (E) Densitometric analysis demonstrating altered protein abundance of ATP7A was performed using ImageJ by normalizing to β-actin. (F–L) mRNA expression of CEBPε (F), ATP7B (G), ATP7A (H), CTR1 (I), CTR2 (J), LOXL2 (K), and AOC2 (L) at day 0, day 2, and day 6 of ATRA-induced NB4 cell differentiation. Values represent mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 relative to the undifferentiated control at day 0 or between multiple time points analyzed by ANOVA. Data represent 3–5 independent experiments. The gray color bar denotes day 0, magenta denotes day 2, and blue denotes day 6 NB4 cells, respectively. Abbreviations: ATRA, all-trans retinoic acid. See also .

Article Snippet: Human myeloid NB4 (Cytion, cat# 300299) and NB4/Luc+ cells (AML-M3, gift from Gabriel Ghiaur’s lab) were cultured in RPMI1640 media supplemented with 10% FBS +2 mM Glutamine +1% Pen-Strep.

Techniques: Cell Differentiation, Western Blot, Quantitative Proteomics, Control, Expressing

Journal: Cell reports

Article Title: ATP7B-maintained copper stores in myeloid progenitors are required for functional maturation of neutrophils

doi: 10.1016/j.celrep.2026.116955

Figure Lengend Snippet: (A) Live images of CopperGREEN in ATRA-induced differentiated NB4 cells (day 6). Scale bar, 10 μm. (B) Cellular copper content at day 0, day 2, and day 6 of ATRA-induced NB4 cell differentiation, measured by ICP-MS, and statistical analysis was performed by ordinary one-way ANOVA. Fg denotes femtogram. (C) CopperGREEN staining in isolated BM progenitors (CD117+) and BM neutrophils reveals an increase in intracellular Cu in BM neutrophils. Representative images of one of the CopperGREEN-stained images is shown from progenitors (CD117+) and neutrophils derived from BM. Scale bar, 10 μm. (D) Fluorescence intensity shows increased Cu levels in BM neutrophils compared to progenitors (CD117+). The data represent two independent experiments. **** p < 0.0001 by t test. (E) CopperGREEN staining was lower in blood neutrophils than in BM neutrophils. Treatment with the Cu chelator TTM (10 μM) reduced intracellular Cu staining in BM neutrophils. BM neutrophils without CopperGREEN were used as a negative control. The data represent three independent experiments. Scale bar, 10 μm.

Article Snippet: Human myeloid NB4 (Cytion, cat# 300299) and NB4/Luc+ cells (AML-M3, gift from Gabriel Ghiaur’s lab) were cultured in RPMI1640 media supplemented with 10% FBS +2 mM Glutamine +1% Pen-Strep.

Techniques: Cell Differentiation, Staining, Isolation, Derivative Assay, Fluorescence, Negative Control

Journal: Frontiers in Immunology

Article Title: Ziyuglycoside II ameliorates chemotherapy-induced neutropenia by promoting neutrophil differentiation and functional recovery via SPI1 and C/EBPϵ transcriptional regulation

doi: 10.3389/fimmu.2026.1771161

Figure Lengend Snippet: Ziyuglycoside II induces differentiation without cell cycle arrest or apoptosis in NB4 cells. (A, B) Dose- and time-dependent antiproliferative effects of ZGSII (0-100 μM, 24–120 h) assessed by CCK-8 assay. (C-F) Flow cytometric analysis demonstrating no significant changes in (C, D) cell cycle progression (propidium iodide staining) or (E, F) apoptosis (Annexin V/PI assay) following 120 h treatment at differentiation-inducing concentrations (5-10 μM). (G-J) Concentration-dependent neutrophilic differentiation evidenced by: (G, H) increased CD11b + cell population (flow cytometry) and (I, J) characteristic morphological maturation in Wright-Giemsa stained NB4 cells (arrows indicate segmented nuclei). Data represent mean ± SD (n=3 independent experiments). * P < 0.05, **** P < 0.0001 versus vehicle control.

Article Snippet: In vitro cell staining : NB4 cells (1×10 6 /mL) were smeared, air-dried, fixed in 4% paraformaldehyde (PFA), and stained (Solution A/B, Servicebio).

Techniques: CCK-8 Assay, Staining, Concentration Assay, Flow Cytometry, Control

Journal: Cell reports

Article Title: ATP7B-maintained copper stores in myeloid progenitors are required for functional maturation of neutrophils

doi: 10.1016/j.celrep.2026.116955

Figure Lengend Snippet: (A) Co-immunostaining of ATP7B (green) with the trans-Golgi marker TGN46 (red) in mouse myelocytes and co-immunostaining of ATP7B (green) with the cis-Golgi marker GM130 (red) in human myeloid NB4 cells. A zoomed view used to demonstrate the ATP7B (green) vesicular pattern. (B) Co-immunostaining of ATP7B (yellow) with TGN46 (cyan), the early endosome marker EEA1 (cyan), the lysosome marker LAMP1 (magenta), and the late endosomal marker Rab8 (magenta) in non-differentiated NB4 cells. There was no significant co-localization of ATP7B with Golgi and endosomal markers, respectively. Scale bar, 5 μm. (C) Live images of CopperGREEN sensor in non-differentiated NB4 cells. DAPI (blue) was used as a nuclear marker. Scale bar, 10 μm. See also – .

Article Snippet: Human NB4 cell line , Cytion , 300299.

Techniques: Immunostaining, Marker

Journal: Cell reports

Article Title: ATP7B-maintained copper stores in myeloid progenitors are required for functional maturation of neutrophils

doi: 10.1016/j.celrep.2026.116955

Figure Lengend Snippet: (A) Schematic representation of ATRA-induced human myeloid NB4 cell differentiation. (B) Western blot analysis reveals that ATP7B protein abundance is significantly reduced and CEBPε is upregulated during ATRA-induced NB4 cell differentiation from day 0 to day 6. β-actin was used as an endogenous loading control. Immunoblotting data represent 4–5 independent experiments. (C) Densitometric analysis demonstrating altered protein abundance of ATP7B and CEBPε was performed using ImageJ by normalizing to β-actin. (D) Western blot analysis reveals that ATP7A protein abundance is upregulated during ATRA-induced NB4 cell differentiation from day 0 to day 6. (E) Densitometric analysis demonstrating altered protein abundance of ATP7A was performed using ImageJ by normalizing to β-actin. (F–L) mRNA expression of CEBPε (F), ATP7B (G), ATP7A (H), CTR1 (I), CTR2 (J), LOXL2 (K), and AOC2 (L) at day 0, day 2, and day 6 of ATRA-induced NB4 cell differentiation. Values represent mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 relative to the undifferentiated control at day 0 or between multiple time points analyzed by ANOVA. Data represent 3–5 independent experiments. The gray color bar denotes day 0, magenta denotes day 2, and blue denotes day 6 NB4 cells, respectively. Abbreviations: ATRA, all-trans retinoic acid. See also .

Article Snippet: Human NB4 cell line , Cytion , 300299.

Techniques: Cell Differentiation, Western Blot, Quantitative Proteomics, Control, Expressing

Journal: Cell reports

Article Title: ATP7B-maintained copper stores in myeloid progenitors are required for functional maturation of neutrophils

doi: 10.1016/j.celrep.2026.116955

Figure Lengend Snippet: (A) Live images of CopperGREEN in ATRA-induced differentiated NB4 cells (day 6). Scale bar, 10 μm. (B) Cellular copper content at day 0, day 2, and day 6 of ATRA-induced NB4 cell differentiation, measured by ICP-MS, and statistical analysis was performed by ordinary one-way ANOVA. Fg denotes femtogram. (C) CopperGREEN staining in isolated BM progenitors (CD117+) and BM neutrophils reveals an increase in intracellular Cu in BM neutrophils. Representative images of one of the CopperGREEN-stained images is shown from progenitors (CD117+) and neutrophils derived from BM. Scale bar, 10 μm. (D) Fluorescence intensity shows increased Cu levels in BM neutrophils compared to progenitors (CD117+). The data represent two independent experiments. **** p < 0.0001 by t test. (E) CopperGREEN staining was lower in blood neutrophils than in BM neutrophils. Treatment with the Cu chelator TTM (10 μM) reduced intracellular Cu staining in BM neutrophils. BM neutrophils without CopperGREEN were used as a negative control. The data represent three independent experiments. Scale bar, 10 μm.

Article Snippet: Human NB4 cell line , Cytion , 300299.

Techniques: Cell Differentiation, Staining, Isolation, Derivative Assay, Fluorescence, Negative Control