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Fig. 1 | Experimental Hematology & Oncology

Fig. 1

From: JOSD2 regulates PKM2 nuclear translocation and reduces acute myeloid leukemia progression

Fig. 1

a mRNA levels of JOSD1 and JOSD2 in peripheral blood mononuclear cells (PBMCs) from primary AML patients compared with normal PBMCs (M0 = 3, M1 = 5, M2 = 5, M3 = 3, M4 = 3, M5 = 5, M6 = 3, normal n = 24). Data are presented as mean ± SD, and were analyzed by using One-way ANOVA. **P < 0.01, ***P < 0.001. b Western blot analysis of JOSD1 and JOSD2 protein expression in PBMCs from health donors and primary AML patients. &represents a band of nonspecific signals. c Cell viability of HL60, U937 and NB4 leukemia cells after JOSD1 and JOSD2 overexpression was measured by CCK-8 assay. Data are presented as mean ± SD, and were analyzed by using the 2-tailed Student t test. **P < 0.01, ***P < 0.001. d Western blot analysis of protein expression as shown in the figure after restoring JOSD2 expression in HL60, U937 and primary AML cells. e Immunopurification and mass spectrometric analysis of JOSD2-interacting proteins in JOSD2-overexpressed HL60 cells. f Co-immunoprecipitation assay showing the interaction between JOSD2 and PKM2 in JOSD2-overexpressed HL60 cells. g JOSD2 and PKM2 were co-located in the cytoplasm of HL60 cells overexpressing JOSD2. h Western blot analysis of PKM2 expression after restoring JOSD2 expression in HL60 cells. i Immunofluorescence assay shows that JOSD2 blocks PKM2 nuclear localization in HL60 cells. Scale bar, 20 μm. j The number of PKM2cells in cytoplasm or nucleus was counted. k The localization of PKM2 in HL60 cells with low JOSD2 expression and high JOSD2 expression was detected by nuclear cytoplasmic separation. Data are presented as mean ± SD, and were analyzed by using the 2-tailed Student t test. ***P < 0.001. l Western blot analysis of c-myc and cyclin D1 expression after restoring JOSD2 expression in HL60 cells. m The effects of JOSD2 on K433 acetylation and phosphorylation of S37 and Y105 of PKM2 were detected by westernblot. n JOSD2 or NC transfected HL60 cells were injected into B-NDG mice (n = 6 in NC grounp, n = 7 in JOSD2 grounp) through the tail vein. The survival was analyzed by Mantel-Cox-log-rank test. ***P < 0.001. o The spleen of leukemia-mice was weighed at day 16. Data are presented as mean ± SD, and were analyzed by using the 2-tailed Student t test. ***P < 0.001. p H&E staining of liver and spleen from mice bearing leukemia at day 16. Scale bar, 200 μm

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