mcur1  (Cell Signaling Technology Inc)

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 is the potential functional target at the 6p23 locus relevant to HAA (A) Population branch statistic (PBS) values of SNPs across the entire human genome. Top 10 loci with their nearest genes are marked by their corresponding colors, with their leading SNPs and PBS values in parentheses. (B) Linkage disequilibrium (LD) structure of the SNPs at the 6p23 locus in the TIB_discovery population. (C) Regional plot of PBS values of SNPs at the 6p23 locus in the discovery stage. (D) Regional plot of probabilities of linked soft selection sweep by diploS/HIC. (E) Association between rs61644582 deletion allele frequencies and the altitude in Asians, Americans, and Africans and their combined populations. (F) Association between rs61644582 genotypes and mRNA levels of eight genes surrounding the 6p23 locus in peripheral blood datasets. (G) Association between rs61644582 genotypes and expression of indicated genes in the peripheral blood of 200 Tibetans (TIB_replication 2).

Article Snippet: To obtain the hematopoiesis-specific Mcur1-cKO mice, Mx1 was used according to the guideline from previous studies., The effect of the Mcur1 (conditional) knockout on mice was validated through Western blotting assays by using the antibodies against MCUR1 (#A08547-1; BosterBio, USA) and β-actin (#60008-1-Ig; Proteintech, China).

Techniques: Functional Assay, Selection, Expressing

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 depletion reduces erythropoiesis in mice (A) The radar plot of the difference in percentages of body weight, spleen weight, and hematological indexes in peripheral blood when comparing Mcur1 -WT and Mcur1 -cKO mice under normoxic or hypoxic conditions. (B) Uniform manifold approximation and projection (UMAP) plot showing the eight cell clusters identified in cKit + cells from BM of Mcur1 -WT and Mcur1 -cKO mice by scRNA-seq. (C–E) The fractions of erythroblasts (C), erythroid progenitors (D), and BFU-E/CFU-E cells (E) in bone marrow (BM). (F and G) The numbers and sizes of the cultured BFU-Es (F) and CFU-Es (G). The representative images of BFU-E/CFU-E are from the 10 th /3 rd day, respectively. (H) UMAP plot showing the reclustering of the HSCs/MPPs from (B). (I) GSEA showing the indicated signatures comparing Mcur1 -WT with Mcur1 -cKO mice under hypoxia. The GSEA was performed by weighted Kolmogorov-Smirnov test. FDR, false discovery rate; ES, enrichment score. Data are shown as the mean ± standard deviation (SD). Data were analyzed by two-tailed t test except where noted otherwise. n.s., not significant. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: To obtain the hematopoiesis-specific Mcur1-cKO mice, Mx1 was used according to the guideline from previous studies., The effect of the Mcur1 (conditional) knockout on mice was validated through Western blotting assays by using the antibodies against MCUR1 (#A08547-1; BosterBio, USA) and β-actin (#60008-1-Ig; Proteintech, China).

Techniques: Cell Culture, Standard Deviation, Two Tailed Test

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 downregulation reduces erythropoiesis in human cord blood CD34 + HSPCs (A) Efficiency of knockdown or reexpression of MCUR1 by western blotting assays. (B–F) MCUR1 knockdown reduces the fraction of benzidine-positive cells (B); CD235a / HBG1 mRNA levels (C); percentages of basophilic (Bas), polychromatophilic (Pol), and orthochromatic (Ort) cells and erythroblasts and erythrocytes (Ery) (D); fraction of erythroblasts (E); and percentages of BFU-E (IL3R − GPA − CD34 + CD36 − ) and CFU-E (IL3R − GPA − CD34 − CD36 + ) populations (F). In (B) and (D), left side shows the representative images. (G and H) MCUR1 knockdown reduces the number and size of colonies formed of BFU-E (G) and CFU-E (H). Representative images of BFU-E/CFU-E at the 14 th /7 th day are shown. (I–K) MCUR1 knockdown leads to reduced cell growth (I), increased cell apoptosis (J), and decreased cell-cycle progression (K). In (I), the analyses between groups were by two-way ANOVA. Data are shown as mean ± SD. Data analyses were conducted by two-tailed t test except where noted otherwise. n.s., not significant. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: To obtain the hematopoiesis-specific Mcur1-cKO mice, Mx1 was used according to the guideline from previous studies., The effect of the Mcur1 (conditional) knockout on mice was validated through Western blotting assays by using the antibodies against MCUR1 (#A08547-1; BosterBio, USA) and β-actin (#60008-1-Ig; Proteintech, China).

Techniques: Knockdown, Western Blot, Two Tailed Test

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 downregulation reduces erythropoiesis by reducing the activation of the mTOR pathway in human cord blood CD34 + HSPCs (A) GSEA showing top enriched hallmark pathways after MCUR1 knockdown in human cord blood CD34 + HSPCs under hypoxia (1% O 2 ) and EPO treatment (3 U/mL) for 7 days. (B) GSEA showing the enrichment of mTOR pathways. (C) MCUR1 knockdown reduces the mTOR activity, and activation of mTOR (by MHY1485 [2 μM]) abolishes the decreased mTOR activity by MCUR1 knockdown. (D) MCUR1 knockdown reduces protein translation rate by polysome profiling assays. OD, optical density; RNP, ribonucleoprotein. (E) The effect of activation of mTOR on the changes in proportions of basophilic (Bas), polychromatophilic (Pol), and orthochromatic (Ort) cells and erythroblasts and erythrocytes (Ery) by MCUR1 knockdown. Left side shows the representative images. (F) The effect of activation of mTOR on the decreased fraction of erythroblasts by MCUR1 knockdown. (G) The effect of activation of mTOR on the decreased percentages of CFU-E by MCUR1 knockdown. (H) The effect of activation of mTOR on the decreased cell growth by MCUR1 knockdown. The analysis of cell growth between groups was by two-way ANOVA. Data are shown as mean ± SD. Data analyses were conducted by two-tailed t test except where noted otherwise. n.s., not significant. ∗∗∗ p < 0.001.

Article Snippet: To obtain the hematopoiesis-specific Mcur1-cKO mice, Mx1 was used according to the guideline from previous studies., The effect of the Mcur1 (conditional) knockout on mice was validated through Western blotting assays by using the antibodies against MCUR1 (#A08547-1; BosterBio, USA) and β-actin (#60008-1-Ig; Proteintech, China).

Techniques: Activation Assay, Knockdown, Activity Assay, Two Tailed Test

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 downregulation reduces activation of the mTOR pathway through the Ca 2+ -CAMMK2-AMPK axis in human cord blood CD34 + HSPCs (A) The effects of MCUR1 knockdown on mitochondrial ([Ca 2+ ] m ; red) and cytoplasmic ([Ca 2+ ] c ; green) Ca 2+ responses in human cord blood CD34 + HSPCs before and after ionomycin exposure (2.5 μM), under hypoxia (1% O 2 ) and EPO treatment (3 U/mL) for 7 days. (B) Knockdown of AMPKa1 induces mTOR activity and abolishes the decreased mTOR activity by MCUR1 knockdown. (C) The effect of AMPKa1 knockdown on the changes in percentages of basophilic (Bas), polychromatophilic (Pol), and orthochromatic (Ort) cells and erythroblasts and erythrocytes (Ery) by MCUR1 knockdown. Left side shows the representative images. (D) The effect of AMPKa1 knockdown on the decreased fraction of erythroblasts by MCUR1 knockdown. (E) The effect of AMPKa1 knockdown on the decreased fraction of CFU-E population by MCUR1 knockdown. (F) The effects of MCUR1 knockdown on the activity of two upstream kinases of AMPKα, LKB1 and CAMKK2. (G) The effects of inhibition of CAMMK2 activity (by BAPTA-AM, an intracellular Ca 2+ chelator; 1 μM) on the increased AMPKα activity and decreased mTOR activity by MCUR1 knockdown. Data are shown as mean ± SD. Data analyses were conducted by two-tailed t test except where noted otherwise. n.s., not significant. ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

Article Snippet: To obtain the hematopoiesis-specific Mcur1-cKO mice, Mx1 was used according to the guideline from previous studies., The effect of the Mcur1 (conditional) knockout on mice was validated through Western blotting assays by using the antibodies against MCUR1 (#A08547-1; BosterBio, USA) and β-actin (#60008-1-Ig; Proteintech, China).

Techniques: Activation Assay, Knockdown, Activity Assay, Inhibition, Two Tailed Test

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: PU.1 preferentially binds to rs61644582 deletion allele and reduces MCUR1 expression to a greater extent compared to C allele (A) A comprehensive map around rs61644582 at the 6p23 locus. The epigenetic modification enrichment profiles in erythroblasts and K562 were derived from the BLUEPRINT and ENCODE projects, respectively. C and /, the C and deletion (del) allele of rs61644582, respectively. (B) The effects of rs61644582 on transcriptional activity of the MCUR1 promoter by luciferase reporter gene assays. E, enhancer; Luc, luciferase gene; P, promoter. (C) The binding affinity of del and C alleles of rs61644582 to PU.1 by EMSA. Competition gradients, which are represented by black triangles, correspond to 5-, 50-, and 100-fold excess of cold probes. (D) The rs61644582 del allele abolishes the binding of a protein complex that is supershifted by the antibody against PU.1 or ELF1. (E) The binding affinity of del and C alleles of rs61644582 to PU.1 or ELF1 in nuclear extract (N.E.) by WEMSA. The protein quantification was determined using ImageJ. (F) Chromatin binding of PU.1 or ELF1 at the rs61644582 site by ChIP-qPCR. (G) PU.1, rather than ELF1, favors binding to the rs61644582 del allele as determined by ChIP-AS-qPCR in HUDEP2 cells, which are heterozygous at rs61644582. (H) PU.1, rather than ELF1, favors binding to the rs61644582 del allele as determined by ChIP-PCR followed by Sanger sequencing. (I) PU.1 knockdown induces MCUR1 expression. (J) Overexpression of PU.1 reduces MCUR1 expression. (K) Association between the mRNA levels of MCUR1 and PU.1 in PBMCs. (L) The stronger inhibitory effect of rs61644582 del allele relative to C allele on luciferase reporter activity was abolished when PU.1 was knocked down. Data are shown as mean ± SD. (M) Association of rs61644582 genotypes with the mRNA levels of MCUR1 and several erythrocyte-related indicators in human cord blood samples of 42 Tibetans (TIB_replication 3). The data in (B), (E–G), and (L) were analyzed using two-tailed t test, while the data in (K) and (M) were analyzed by linear regression analysis. n.s., not significant. ∗∗∗ p < 0.001.

Article Snippet: To obtain the hematopoiesis-specific Mcur1-cKO mice, Mx1 was used according to the guideline from previous studies., The effect of the Mcur1 (conditional) knockout on mice was validated through Western blotting assays by using the antibodies against MCUR1 (#A08547-1; BosterBio, USA) and β-actin (#60008-1-Ig; Proteintech, China).

Techniques: Expressing, Modification, Derivative Assay, Activity Assay, Luciferase, Binding Assay, ChIP-qPCR, Sequencing, Knockdown, Over Expression, Two Tailed Test

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: A model of the allele-specific transcriptional regulation of MCUR1 by rs61644582 and the function and underlying mechanism of MCUR1 in regulating erythropoiesis The positively selected beneficial rs61644582 del allele has a stronger binding ability to the transcription factor PU.1 relative to the C allele, therefore resulting in relatively lower expression of MCUR1 . Downregulation of MCUR1 reduces the mitochondrial Ca 2+ uptake and then concomitantly increases the cytosolic Ca 2+ levels, thereby attenuating erythropoiesis via the CAMKK2-AMPK-mTOR axis.

Article Snippet: To obtain the hematopoiesis-specific Mcur1-cKO mice, Mx1 was used according to the guideline from previous studies., The effect of the Mcur1 (conditional) knockout on mice was validated through Western blotting assays by using the antibodies against MCUR1 (#A08547-1; BosterBio, USA) and β-actin (#60008-1-Ig; Proteintech, China).

Techniques: Binding Assay, Expressing

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet:

Article Snippet: To obtain the hematopoiesis-specific Mcur1-cKO mice, Mx1 was used according to the guideline from previous studies., The effect of the Mcur1 (conditional) knockout on mice was validated through Western blotting assays by using the antibodies against MCUR1 (#A08547-1; BosterBio, USA) and β-actin (#60008-1-Ig; Proteintech, China).

Techniques: Recombinant, Modification, Protease Inhibitor, Luminescence Assay, DNA Extraction, Multiplex Assay, Reporter Assay, Reverse Transcription, SYBR Green Assay, Electrophoresis, Mobility Shift, Transfection, Chromatin Immunoprecipitation, Mutagenesis, Enzyme-linked Immunosorbent Assay, Purification, Sequencing, RNA Sequencing, Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Plasmid Preparation, Software

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 is the potential functional target at the 6p23 locus relevant to HAA (A) Population branch statistic (PBS) values of SNPs across the entire human genome. Top 10 loci with their nearest genes are marked by their corresponding colors, with their leading SNPs and PBS values in parentheses. (B) Linkage disequilibrium (LD) structure of the SNPs at the 6p23 locus in the TIB_discovery population. (C) Regional plot of PBS values of SNPs at the 6p23 locus in the discovery stage. (D) Regional plot of probabilities of linked soft selection sweep by diploS/HIC. (E) Association between rs61644582 deletion allele frequencies and the altitude in Asians, Americans, and Africans and their combined populations. (F) Association between rs61644582 genotypes and mRNA levels of eight genes surrounding the 6p23 locus in peripheral blood datasets. (G) Association between rs61644582 genotypes and expression of indicated genes in the peripheral blood of 200 Tibetans (TIB_replication 2).

Article Snippet: Further, the Mcur1 conditional KO ( Mcur1-cKO ) mouse model was constructed on the C57BL/6 background in Shanghai Model Organisms Center (China).

Techniques: Functional Assay, Selection, Expressing

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 depletion reduces erythropoiesis in mice (A) The radar plot of the difference in percentages of body weight, spleen weight, and hematological indexes in peripheral blood when comparing Mcur1 -WT and Mcur1 -cKO mice under normoxic or hypoxic conditions. (B) Uniform manifold approximation and projection (UMAP) plot showing the eight cell clusters identified in cKit + cells from BM of Mcur1 -WT and Mcur1 -cKO mice by scRNA-seq. (C–E) The fractions of erythroblasts (C), erythroid progenitors (D), and BFU-E/CFU-E cells (E) in bone marrow (BM). (F and G) The numbers and sizes of the cultured BFU-Es (F) and CFU-Es (G). The representative images of BFU-E/CFU-E are from the 10 th /3 rd day, respectively. (H) UMAP plot showing the reclustering of the HSCs/MPPs from (B). (I) GSEA showing the indicated signatures comparing Mcur1 -WT with Mcur1 -cKO mice under hypoxia. The GSEA was performed by weighted Kolmogorov-Smirnov test. FDR, false discovery rate; ES, enrichment score. Data are shown as the mean ± standard deviation (SD). Data were analyzed by two-tailed t test except where noted otherwise. n.s., not significant. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: Further, the Mcur1 conditional KO ( Mcur1-cKO ) mouse model was constructed on the C57BL/6 background in Shanghai Model Organisms Center (China).

Techniques: Cell Culture, Standard Deviation, Two Tailed Test

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 downregulation reduces erythropoiesis in human cord blood CD34 + HSPCs (A) Efficiency of knockdown or reexpression of MCUR1 by western blotting assays. (B–F) MCUR1 knockdown reduces the fraction of benzidine-positive cells (B); CD235a / HBG1 mRNA levels (C); percentages of basophilic (Bas), polychromatophilic (Pol), and orthochromatic (Ort) cells and erythroblasts and erythrocytes (Ery) (D); fraction of erythroblasts (E); and percentages of BFU-E (IL3R − GPA − CD34 + CD36 − ) and CFU-E (IL3R − GPA − CD34 − CD36 + ) populations (F). In (B) and (D), left side shows the representative images. (G and H) MCUR1 knockdown reduces the number and size of colonies formed of BFU-E (G) and CFU-E (H). Representative images of BFU-E/CFU-E at the 14 th /7 th day are shown. (I–K) MCUR1 knockdown leads to reduced cell growth (I), increased cell apoptosis (J), and decreased cell-cycle progression (K). In (I), the analyses between groups were by two-way ANOVA. Data are shown as mean ± SD. Data analyses were conducted by two-tailed t test except where noted otherwise. n.s., not significant. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: Further, the Mcur1 conditional KO ( Mcur1-cKO ) mouse model was constructed on the C57BL/6 background in Shanghai Model Organisms Center (China).

Techniques: Knockdown, Western Blot, Two Tailed Test

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 downregulation reduces erythropoiesis by reducing the activation of the mTOR pathway in human cord blood CD34 + HSPCs (A) GSEA showing top enriched hallmark pathways after MCUR1 knockdown in human cord blood CD34 + HSPCs under hypoxia (1% O 2 ) and EPO treatment (3 U/mL) for 7 days. (B) GSEA showing the enrichment of mTOR pathways. (C) MCUR1 knockdown reduces the mTOR activity, and activation of mTOR (by MHY1485 [2 μM]) abolishes the decreased mTOR activity by MCUR1 knockdown. (D) MCUR1 knockdown reduces protein translation rate by polysome profiling assays. OD, optical density; RNP, ribonucleoprotein. (E) The effect of activation of mTOR on the changes in proportions of basophilic (Bas), polychromatophilic (Pol), and orthochromatic (Ort) cells and erythroblasts and erythrocytes (Ery) by MCUR1 knockdown. Left side shows the representative images. (F) The effect of activation of mTOR on the decreased fraction of erythroblasts by MCUR1 knockdown. (G) The effect of activation of mTOR on the decreased percentages of CFU-E by MCUR1 knockdown. (H) The effect of activation of mTOR on the decreased cell growth by MCUR1 knockdown. The analysis of cell growth between groups was by two-way ANOVA. Data are shown as mean ± SD. Data analyses were conducted by two-tailed t test except where noted otherwise. n.s., not significant. ∗∗∗ p < 0.001.

Article Snippet: Further, the Mcur1 conditional KO ( Mcur1-cKO ) mouse model was constructed on the C57BL/6 background in Shanghai Model Organisms Center (China).

Techniques: Activation Assay, Knockdown, Activity Assay, Two Tailed Test

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 downregulation reduces activation of the mTOR pathway through the Ca 2+ -CAMMK2-AMPK axis in human cord blood CD34 + HSPCs (A) The effects of MCUR1 knockdown on mitochondrial ([Ca 2+ ] m ; red) and cytoplasmic ([Ca 2+ ] c ; green) Ca 2+ responses in human cord blood CD34 + HSPCs before and after ionomycin exposure (2.5 μM), under hypoxia (1% O 2 ) and EPO treatment (3 U/mL) for 7 days. (B) Knockdown of AMPKa1 induces mTOR activity and abolishes the decreased mTOR activity by MCUR1 knockdown. (C) The effect of AMPKa1 knockdown on the changes in percentages of basophilic (Bas), polychromatophilic (Pol), and orthochromatic (Ort) cells and erythroblasts and erythrocytes (Ery) by MCUR1 knockdown. Left side shows the representative images. (D) The effect of AMPKa1 knockdown on the decreased fraction of erythroblasts by MCUR1 knockdown. (E) The effect of AMPKa1 knockdown on the decreased fraction of CFU-E population by MCUR1 knockdown. (F) The effects of MCUR1 knockdown on the activity of two upstream kinases of AMPKα, LKB1 and CAMKK2. (G) The effects of inhibition of CAMMK2 activity (by BAPTA-AM, an intracellular Ca 2+ chelator; 1 μM) on the increased AMPKα activity and decreased mTOR activity by MCUR1 knockdown. Data are shown as mean ± SD. Data analyses were conducted by two-tailed t test except where noted otherwise. n.s., not significant. ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

Article Snippet: Further, the Mcur1 conditional KO ( Mcur1-cKO ) mouse model was constructed on the C57BL/6 background in Shanghai Model Organisms Center (China).

Techniques: Activation Assay, Knockdown, Activity Assay, Inhibition, Two Tailed Test

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: PU.1 preferentially binds to rs61644582 deletion allele and reduces MCUR1 expression to a greater extent compared to C allele (A) A comprehensive map around rs61644582 at the 6p23 locus. The epigenetic modification enrichment profiles in erythroblasts and K562 were derived from the BLUEPRINT and ENCODE projects, respectively. C and /, the C and deletion (del) allele of rs61644582, respectively. (B) The effects of rs61644582 on transcriptional activity of the MCUR1 promoter by luciferase reporter gene assays. E, enhancer; Luc, luciferase gene; P, promoter. (C) The binding affinity of del and C alleles of rs61644582 to PU.1 by EMSA. Competition gradients, which are represented by black triangles, correspond to 5-, 50-, and 100-fold excess of cold probes. (D) The rs61644582 del allele abolishes the binding of a protein complex that is supershifted by the antibody against PU.1 or ELF1. (E) The binding affinity of del and C alleles of rs61644582 to PU.1 or ELF1 in nuclear extract (N.E.) by WEMSA. The protein quantification was determined using ImageJ. (F) Chromatin binding of PU.1 or ELF1 at the rs61644582 site by ChIP-qPCR. (G) PU.1, rather than ELF1, favors binding to the rs61644582 del allele as determined by ChIP-AS-qPCR in HUDEP2 cells, which are heterozygous at rs61644582. (H) PU.1, rather than ELF1, favors binding to the rs61644582 del allele as determined by ChIP-PCR followed by Sanger sequencing. (I) PU.1 knockdown induces MCUR1 expression. (J) Overexpression of PU.1 reduces MCUR1 expression. (K) Association between the mRNA levels of MCUR1 and PU.1 in PBMCs. (L) The stronger inhibitory effect of rs61644582 del allele relative to C allele on luciferase reporter activity was abolished when PU.1 was knocked down. Data are shown as mean ± SD. (M) Association of rs61644582 genotypes with the mRNA levels of MCUR1 and several erythrocyte-related indicators in human cord blood samples of 42 Tibetans (TIB_replication 3). The data in (B), (E–G), and (L) were analyzed using two-tailed t test, while the data in (K) and (M) were analyzed by linear regression analysis. n.s., not significant. ∗∗∗ p < 0.001.

Article Snippet: Further, the Mcur1 conditional KO ( Mcur1-cKO ) mouse model was constructed on the C57BL/6 background in Shanghai Model Organisms Center (China).

Techniques: Expressing, Modification, Derivative Assay, Activity Assay, Luciferase, Binding Assay, ChIP-qPCR, Sequencing, Knockdown, Over Expression, Two Tailed Test

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: A model of the allele-specific transcriptional regulation of MCUR1 by rs61644582 and the function and underlying mechanism of MCUR1 in regulating erythropoiesis The positively selected beneficial rs61644582 del allele has a stronger binding ability to the transcription factor PU.1 relative to the C allele, therefore resulting in relatively lower expression of MCUR1 . Downregulation of MCUR1 reduces the mitochondrial Ca 2+ uptake and then concomitantly increases the cytosolic Ca 2+ levels, thereby attenuating erythropoiesis via the CAMKK2-AMPK-mTOR axis.

Article Snippet: Further, the Mcur1 conditional KO ( Mcur1-cKO ) mouse model was constructed on the C57BL/6 background in Shanghai Model Organisms Center (China).

Techniques: Binding Assay, Expressing

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet:

Article Snippet: Further, the Mcur1 conditional KO ( Mcur1-cKO ) mouse model was constructed on the C57BL/6 background in Shanghai Model Organisms Center (China).

Techniques: Recombinant, Modification, Protease Inhibitor, Luminescence Assay, DNA Extraction, Multiplex Assay, Reporter Assay, Reverse Transcription, SYBR Green Assay, Electrophoresis, Mobility Shift, Transfection, Chromatin Immunoprecipitation, Mutagenesis, Enzyme-linked Immunosorbent Assay, Purification, Sequencing, RNA Sequencing, Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Plasmid Preparation, Software

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet: MCUR1 downregulation reduces activation of the mTOR pathway through the Ca 2+ -CAMMK2-AMPK axis in human cord blood CD34 + HSPCs (A) The effects of MCUR1 knockdown on mitochondrial ([Ca 2+ ] m ; red) and cytoplasmic ([Ca 2+ ] c ; green) Ca 2+ responses in human cord blood CD34 + HSPCs before and after ionomycin exposure (2.5 μM), under hypoxia (1% O 2 ) and EPO treatment (3 U/mL) for 7 days. (B) Knockdown of AMPKa1 induces mTOR activity and abolishes the decreased mTOR activity by MCUR1 knockdown. (C) The effect of AMPKa1 knockdown on the changes in percentages of basophilic (Bas), polychromatophilic (Pol), and orthochromatic (Ort) cells and erythroblasts and erythrocytes (Ery) by MCUR1 knockdown. Left side shows the representative images. (D) The effect of AMPKa1 knockdown on the decreased fraction of erythroblasts by MCUR1 knockdown. (E) The effect of AMPKa1 knockdown on the decreased fraction of CFU-E population by MCUR1 knockdown. (F) The effects of MCUR1 knockdown on the activity of two upstream kinases of AMPKα, LKB1 and CAMKK2. (G) The effects of inhibition of CAMMK2 activity (by BAPTA-AM, an intracellular Ca 2+ chelator; 1 μM) on the increased AMPKα activity and decreased mTOR activity by MCUR1 knockdown. Data are shown as mean ± SD. Data analyses were conducted by two-tailed t test except where noted otherwise. n.s., not significant. ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

Article Snippet: The small interfering RNAs (siRNAs) targeting MCUR1 , TSC1 , TSC2 , AMPKa1 , CAMKK2 , PU.1 and a non-targeting control siRNA were synthesized by RiboBio (Guangzhou City, China).

Techniques: Activation Assay, Knockdown, Activity Assay, Inhibition, Two Tailed Test