Fig. 6: SYNCRIP controls HSC polarity and symmetric vs. asymmetric division. | Nature Communications

Fig. 6: SYNCRIP controls HSC polarity and symmetric vs. asymmetric division.

From: RNA binding protein SYNCRIP maintains proteostasis and self-renewal of hematopoietic stem and progenitor cells

Fig. 6

A Representative images of immunofluorescence (IF) staining of SYNCRIP, CDC42 and TUBULIN in hematopoietic stem cells (HSCs). B–D Quantification of normalized IF intensity SYNCRIP (Syncripf/f n = 379, SyncripΔ/Δ n = 300) (B) and CDC42 (Syncripf/f n = 133, SyncripΔ/Δ n = 129) (C) and TUBULIN (Syncripf/f n = 2482, SyncripΔ/Δ n = 1703) (D). All p < 0.0001. E Percentage of polarized vs. unpolarized HSCs based on TUBULIN (n = 4-5/genotype; Syncripf/f n = 99 and SyncripΔ/Δ n = 175). F Quantification of normalized IF NUMB intensity in HSCs. (Syncripf/f n = 1319 and SyncripΔ/Δ n = 929, p < 0.0001). G Representative images of paired NUMB daughter assay of HSCs. H Percentage of doublet cells in each type of cell division (n = 4-5/genotype; Syncripf/f n = 108 and SyncripΔ/Δ n = 52). I Representative images of IF staining of TUBLIN, LAMP1 and DAPI in HSCs. J Quantification of normalized IF LAMP1 intensity in HSCs. (Syncripf/f n = 566 and SyncripΔ/Δ n = 566, p < 0.0001). K Representative images of paired LAMP1 daughter assay of HSCs. L Percentages of doublet cells in each type of cell division (n = 4-5 each genotype; Syncripf/f n = 110 and SyncripΔ/Δ n = 165). M Representative images of paired TMI daughter assay of HSCs. N Percentages of doublet cells in each type of cell division (n = 4-5 each genotype; Syncripf/f n = 71 and SyncripΔ/Δ n = 38). O Representative images of paired TMI and LAMP1 daughter assay of HSCs. P Percentages of asymmetric divided cells. (n = 4-5 each genotype; Syncripf/f n = 10 and SyncripΔ/Δ n = 6). Q, R Quantification of normalized IF intensity of TMI in doublet cells symmetric high LAMP1 Syncripf/f n = 40 and SyncripΔ/Δ n = 8, p = 0.018 (Q) and symmetric low LAMP1 Syncripf/f n = 56 and SyncripΔ/Δ n = 48, p = 0.022 (R). S Normalized colony numbers 1st, 2nd and 3rd plating of Syncripf/f and SyncripΔ/Δ LSK cells transduced with empty vector (EV) or expressing CDC42 (CDC42-OV) (n = 7/condition). T Percentages of polarized vs. unpolarized cells based on TUBULIN (Syncripf/f-EV n = 20; SyncripΔ/Δ-EV n = 29; Syncrip f/f CDC42-OV n = 22; and SyncripΔ/Δ CDC42-OV n = 19). Scale bars 5μm. Source data are provided as Source Data File. All data represent mean ± s.e.m. p values were calculated by two-tailed t-test. *p < 0.05,**p < 0.01, ***p < 0.001 and ns: not significant.

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