Research Paper Volume 13, Issue 10 pp 13644—13662

microRNA-1246-containing extracellular vesicles from acute myeloid leukemia cells promote the survival of leukemia stem cells via the LRIG1-meditated STAT3 pathway

miR-1246 targets LRIG1 and activates the STAT3 pathway, facilitating cell survival and suppressing differentiation of LSCs. (A) Expression of the STAT3 pathway-related proteins (phosphorylated JAK2 and phosphorylated STAT3) in LSCs co-cultured with AML cell-derived EVs measured by western blot analysis. (B) Expression of STAT3 pathway-related proteins (phosphorylated JAK2 and phosphorylated STAT3) in LSCs after over-expression of miR-1246 and inhibition of LRIG1 alone or in combination demonstrated by western blot analysis. (C) Expression of STAT3 pathway-related proteins (phosphorylated JAK2 and phosphorylated STAT3) in LSCs after exposure to the JAK2 inhibitor AZD1480 determined by western blot analysis. (D) LSCs viability after exposure to the JAK2 inhibitor AZD1480 detected by MTS assay. (E) Colony formation of LSCs after exposure to the JAK2 inhibitor AZD1480 evaluated by colony formation assay. (F) LSCs apoptosis after exposure to the JAK2 inhibitor AZD1480 identified by flow cytometry. (G) Differentiation of LSCs after exposure to the JAK2 inhibitor AZD1480 determined by differentiation assay. * p vs. LSCs co-cultured with PBS, EVs-inhibitor-NC, mimic-NC, si-NC, LRIG1-NC, miR-1246 mimic + LRIG1-NC or DMSO. Data in the figures are all measurement data, expressed as mean ± standard deviation. Data among multiple groups were analyzed by one-way ANOVA. The experiments were repeated in triplicate.

Figure 6. miR-1246 targets LRIG1 and activates the STAT3 pathway, facilitating cell survival and suppressing differentiation of LSCs. (A) Expression of the STAT3 pathway-related proteins (phosphorylated JAK2 and phosphorylated STAT3) in LSCs co-cultured with AML cell-derived EVs measured by western blot analysis. (B) Expression of STAT3 pathway-related proteins (phosphorylated JAK2 and phosphorylated STAT3) in LSCs after over-expression of miR-1246 and inhibition of LRIG1 alone or in combination demonstrated by western blot analysis. (C) Expression of STAT3 pathway-related proteins (phosphorylated JAK2 and phosphorylated STAT3) in LSCs after exposure to the JAK2 inhibitor AZD1480 determined by western blot analysis. (D) LSCs viability after exposure to the JAK2 inhibitor AZD1480 detected by MTS assay. (E) Colony formation of LSCs after exposure to the JAK2 inhibitor AZD1480 evaluated by colony formation assay. (F) LSCs apoptosis after exposure to the JAK2 inhibitor AZD1480 identified by flow cytometry. (G) Differentiation of LSCs after exposure to the JAK2 inhibitor AZD1480 determined by differentiation assay. * p < 0.05 vs. LSCs co-cultured with PBS, EVs-inhibitor-NC, mimic-NC, si-NC, LRIG1-NC, miR-1246 mimic + LRIG1-NC or DMSO. Data in the figures are all measurement data, expressed as mean ± standard deviation. Data among multiple groups were analyzed by one-way ANOVA. The experiments were repeated in triplicate.