Fig. 5: Effect of 9p ploidy and JAK2 mutational status on PD-L1, OCT4, and NANOG expression and their role in CD34+ cells clonogenic potential. | Leukemia

Fig. 5: Effect of 9p ploidy and JAK2 mutational status on PD-L1, OCT4, and NANOG expression and their role in CD34+ cells clonogenic potential.

From: Chromosome 9p trisomy increases stem cells clonogenic potential and fosters T-cell exhaustion in JAK2-mutant myeloproliferative neoplasms

Fig. 5

A Experimental workflow of genomic/transcriptomic-coupled characterization of single-cell derived colonies by means of ddPCR. Evaluation of CD274 (B, C), POU5F1 (D, E), and NANOG (F, G) expression at the transcriptional level in single-cell derived colonies by ddPCR. In (B, D, and F) colonies (n = 54) were split according to the number of JAK2 mutated alleles and 9p ploidy status. The light blue area highlights +9p colonies. On the other hand, colonies were split according to 9p ploidy status in (C, E, and G). Results are represented as mean + SEM. H–J POU5F1 and NANOG silencing in CD34+ cells from HOM (n = 4) and +9p (n = 6) patients. H Ratio between the total number of colonies grown in methylcellulose-based clonogenic assays in OCT4siRNA+NANOGsiRNA and NTsiRNA samples, split between HOM and +9p patients. Results are represented as mean + SEM. I Ratio between the number of colonies grown in methylcellulose-based clonogenic assays in OCT4siRNA + NANOGsiRNA and NTsiRNA samples, split according to the different Colony-Forming Units (CFUs) in HOM and +9p patients. Results are represented as mean + SEM. J Barplot depicting the frequency of diploid and +9p colonies in +9p patients (n = 6) after POU5F1 and NANOG silencing or NTsiRNA nucleofection. RQ: relative quantity; 1 mut: one JAK2V617F-mutated allele; 2 mut: two JAK2V617F-mutated alleles; 2n: diploid colonies; 3n: +9p colonies; CFU-E/BFU-E: Colony-Forming Unit—Erythroid/Burst-Forming Unit—Erythroid; CFU-GM: Colony-Forming Unit—Granulocyte/Macrophage; CFU-G = Colony-Forming Unit—Granulocyte; CFU-M = Colony-Forming Unit—Macrophage; CFU-GEMM = Colony-Forming Unit—Granulocyte/Erythrocyte/Macrophage/Megakaryocyte; HOM: JAK2-mutated homozygous patients; +9p: JAK2-mutated patients with 9p trisomy; OCT4 + NANOG: POU5F1 siRNA + NANOG siRNA; NT siRNA: Non Targeting siRNA. *p < 0.05; **p < 0.01. A Was created with Biorender.com.

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