Journal: Aging Cell
Article Title: Periodic Therapeutic Phlebotomy Mitigates Systemic Aging Phenotypes by Promoting Bone Marrow Function
doi: 10.1111/acel.70400
Figure Lengend Snippet: PTP promotes the transformation of hematopoiesis to a youthful state by improving the hematopoietic microenvironment in the bone marrow. (A) Representative SA‐β‐gal staining of rat bone marrow (scale bars, 200 μm) (B) Quantification of SA‐β‐gal + cells per area, n = 3/group. (C) TGF‐β1 (pg/mL) and TPO (pg/mL) in rat bone marrow, n = 3/group. (D–N) The proportion of lin − cell subsets (HSC, LT‐HSC, ST‐HSC, MPP1, MPP5, MPP2, MPP3, CMP, MPP4, CLP, Pre GM, GMP, Pre MegE, MEP, MKP) in mice bone marrow cells, n = 3–4/group. Statistical analysis: One‐way ANOVA and Bonferroni post‐test. (O) Multiple cytokine profiling quantification of SASP proteins in mice bone marrow, log2‐transformed fold change in mean fluorescence intensity (MFI) compared to the average of D‐gal group, n = 6/group. (P) Representative examples and (Q) quantification of SEC (CD31 low Sca‐1 low ) and AEC (CD31 hi Sca‐1 hi ) in ECs from rat bone marrow and endosteum was detected by flow cytometry, n = 3/group. (R) The third generation of rat bone marrow‐derived BMSCs were cultured for 8 days. Cell viability was measured by MTT assay, n = 5/group. (S–T) Western blot of P16, P21, P27, and P63 expression in rat bone marrow‐derived BMSCs, n = 4/group. (U) Cell cycle analysis of rat bone marrow‐derived BMSCs by flow cytometry, n = 3/group. (V) Quantification of cell cycle phase distribution, n = 3/group. (W) Apoptosis of the third generation of rat bone marrow‐derived BMSCs using Annexin V‐FITC/PI KIT combined with flow cytometry, n = 3/group. Statistical analysis: One‐way ANOVA, Bonferroni post‐test for (D–N) and Tukey post‐test for the others, Data are mean ± SD; error bars denote 95% CI; * p < 0.05, ** p < 0.01, *** p < 0.001.
Article Snippet: The concentration of Klotho (Elabscience, E‐EL‐R2580c), Taurine (Absin, abs580222), TGF‐β1 (Multi Sciences, EK981), TPO (Fine Test, ER0202) was measured in accordance with the protocols outlined in the respective reagent kit manuals.
Techniques: Transformation Assay, Staining, Fluorescence, Flow Cytometry, Derivative Assay, Cell Culture, MTT Assay, Western Blot, Expressing, Cell Cycle Assay
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