Fig 1: BM-iPSC-derived bone marrow (BM) milieu supports human hematopoietic cells ex vivo(A) Schema for synthetic RNA-based reprogramming using pluripotent transcripts POU5F1-SOX2-KLF4, GLIS1. Scale bar, 100 µM. Two BM-MSC samples (2 biological replicates) reprogrammed to form 13 BM-iPSC lines.(B) H&E staining of BM-iPSC-derived teratomas (5 NSG mice per i-niche sample) representing the 3 embryonic lineages. Scale bar, 100 µM.(C) Scatterplot showing comparable gene expression between primary BM mesenchymal stem cells (BM-MSC) and BM-iPSC derived MSC (iMSC). The genes profiled include MSC-specific genes IGF1, HGF, VIM, KITLG, PTPRC, PIGS, MMP2, ICAM1, COL1A1, VEGFA, TGFB3, SLC17A5, GTF3A, IL1B, NES, EGF, ITGB1, ANXA5, CSF2, CTNNB1, NUDT6, FUT1, BDNF, BGLAP, FGF22, LIF, ZFP42, SOX2, POU5F1, PROM1, CD44, MCAM, ITGA6, COL9A1, PDGFRB, NT5E, ITGAV, COL2A1, ERBB2, THY1, VCAM1, and ANPEP.(D) GDF6, BMP6, and RUNX2 expression in i-MSC-derived cartilage/chondrocytes, bone/osteoblasts, and fat/adipocytes cells (iC, iO, and iA). Immunohistochemical staining (2 technical replicates) demonstrating safranin O, alizarin red and oil red O staining in iC, iO, and iA, respectively. Scale bar, 100 µM.(E) mRNA expression relative to HKG (housekeeping genes: ACTB, B2M, GAPDH, HPRT1, and RPLP0) in iANG containing representative vascular cells such as CD31+ endothelial cells and CD31- perivascular cells in known proportions (Figure S2).(E) Gene expression has been normalized with respect to HKG (housekeeping genes: ACTB, B2M, GAPDH, HPRT1, and RPLP0) and the fold change expression between CD31+ endothelial cells/CD31- perivascular cells has been plotted. CD31+ cells express endothelial-relevant markers such as APOE, OCLN, ADAM17, and VCAM1, whereas CD31- cells express perivascular markers such as ANXA5, ITGB1, HIF1A, and COL18A1.(F) Cell counts of CD45+ hematopoietic cells (3 biological replicates) extracted from non-malignant human BM and co-cultured on iMSC, iANG versus in niche-free suspension cultures over 7 days.
Fig 2: SETD2 deficiency is associated with decreased turnover of mutant HTT protein and increased cellular toxicity.a, b HeLa cells were transfected with a control non-targeting siRNAs pool or SETD2 siRNAs pool for 48 h and thereafter transfected with the wild-type HTT16Q-CFP or disease-associated mutant mHTT94Q-CFP expression vectors for 24 h prior to protein aggregates assessment. Quantification of aggregates per cell is depicted in panel b. c Immunoblot analysis of HTT16Q and mHTT94Q expression levels in cells, under identical conditions as described above, using an antibody targeting polyQ. d Cell death analysis of HeLa cells transfected with wild-type HTT16Q or disease-associated mutant mHTT94Q plasmid with or without siRNA SETD2 (a non-targeting siRNAs pool was used as a control). Propidium iodide is shown on the Y axis and ANXA5/annexin-V-APC on the X axis. Bars display the mean of at least three independent experiments (a–d, n = 3; e, n = 5), error bars represent SEM; *P < 0.05, **P < 0.01, ***P < 0.001.
Supplier Page from BioLegend for APC Annexin V Apoptosis Detection Kit with PI