Extended Data Fig. 6: Characterization of ER stress and UPR activation in Sel1l-KO HSCs.
a, Transmission electron microscopic analysis of the ER and mitochondria (M) in HSCs from Control (Ctrl) or Sel1lVav-KO mice. Scale bar, 1 μm. b, ER-tracker staining in Ctrl and Sel1lMx1 HSCs. n = 3. c, WB analysis of HSCs from 8-week-old control and Sel1lVav-KO mice. d, RT-PCR analysis of UPR and ERAD in 8-week-old control or Sel1lVav-KO HSCs. Ctrl: n = 4; Sel1lVav: n = 3. e, f, RT-PCR analysis of Xbp1s (e) and Bip (f) expression in c-Kit-enriched BM cells from Ctrl and Sel1lVav-KO mice treated with vehicle or TUDCA (TUDCA-2: 200 mg/kg, TUDCA-5: 500 mg/kg). Vehicle: n = 2; TUDCA: n = 3. g, Frequency of HSCs in vehicle- or TUDCA (200 mg/kg)-treated Ctrl and Sel1lVav mice. n = 2 for vehicle-treated control group and n = 3 for the other 3 groups. h, i, Percentage of control and Sel1lMx1 donor-derived PBMC (h) and HSCs (i) in recipient mice at indicated time after vehicle or TUDCA treatment (daily i.p. injection) starting from the first poly(I:C) injection (4 weeks after donor-reconstitution). HSCs were analysed at week 5. Ctrl: n = 2; Sel1lMx1: n = 3. The Vehicle groups and donor chimerism of n=3 randomly sampled mice from control or Sel1lMx1 donor group before poly(I:C) injection (left panel of h) is the same as in Extended Data Figs. 6k,l, 7 and 8. j, RT-PCR analysis of Xbp1s expression in LSK cells from 4-PBA treated mice. n = 2. k, l, Percentage of Ctrl and Sel1lMx1 donor-derived PBMC (k) and HSCs (l) in recipient mice at indicated time after vehicle or 4-PBA treatment (in drinking water) starting from the first poly(I:C) injection. Ctrl: n = 2; Sel1lMx1: n = 3. Results are shown as mean ± s.d. Two-tailed Student’s t-tests (b, d, j) or two-way ANOVA with Bonferroni test (e-i, k, l) were used to calculate P values. ns, not significant. Statistical information and unprocessed blots are provided as source data.
