Journal: Nature Communications

Article Title: Expansion of interferon inducible gene pool via USP18 inhibition promotes cancer cell pyroptosis

doi: 10.1038/s41467-022-35348-5

Figure Lengend Snippet: a Western blot of WT, PLK2 − / − , USP18 − / − , and USP18 − / − PLK2 − / − THP-1 cells treated with or without IFNα (1000 U/ml) for 48 h with indicated antibodies. b Percentage of live cells in ( a ) ( n = 3 independent samples each). p -value was determined by one-way ANOVA test. c Analysis of ICD markers (LDH release and ecto-CRT) in ( b ) ( n = 3 independent samples each). p -value was determined by one-way ANOVA test. d Western blot of PLK2 − / − THP-1 cells infected with Mock or PLK2 after two days of blasticidin selection using the indicated antibodies. e; Percentage of live cells in ( d ) ( n = 3 independent samples each). p -value was determined by two-tailed Student t -test. f Analysis of ICD markers (LDH release and ecto-CRT) and IL-1β secretions in ( d ) ( n = 3 independent samples each). p -value was determined by two-tailed Student t -test. g ; THP-1 and Kasumi-1 cells were transduced with MIP, MIP-PLK2 WT, or MIP-PLK2 enzyme activity mutants. Two days after puromycin selection, cell lysates were analyzed by western blot with indicated antibodies. h Ectopic PLK2 expression induces cell death in several AML cell lines independent of its kinase activity. THP-1, Kasumi-1, and MV4-11 cells were infected with MIP, MIP-PLK2 WT, or MIP-PLK2 enzyme activity mutants. Two days after puromycin selection, live cells were analyzed by FACS ( n = 3 independent samples each). p -value was determined by one-way ANOVA test. i THP-1 or OCI-AML3 cells were treated with DMSO or TC-S7005 (final 10 µM) for 48 h, then cell lysates were analyzed. j THP-1 or OCI-AML3 cells were treated with DMSO or TC-S7005 for indicated times, then PI (%) and Annexin V + (%) were analyzed by FACS ( n = 3 independent samples each). p -value was determined by one-way ANOVA test. k Two different PDX AML cells were cultured with different dose of TC-S7005, and analyzed cell growth ( n = 3 independent samples each conditions). All data represent mean ± s.d, except where indicated. n.s. not statistically significant. p -value was determined by one-way ANOVA test. Source data are provided as a Source Data file.

Article Snippet: THP-1 (ATCC), MV4-11 (ATCC), MOLM13 (ATCC), OCI-AML3 (ATCC), Kasumi-1 (ATCC), B16F10 (ATCC) and MC38 (NCI) cells were grown in RPMI media supplemented with glutamine, penicillin/streptomycin, and 10% FBS.

Techniques: Western Blot, Infection, Selection, Two Tailed Test, Transduction, Activity Assay, Expressing, Cell Culture

Journal: Journal of Translational Medicine

Article Title: Downregulation of SMIM3 inhibits growth of leukemia via PI3K-AKT signaling pathway and correlates with prognosis of adult acute myeloid leukemia with normal karyotype

doi: 10.1186/s12967-022-03831-8

Figure Lengend Snippet: Downregulation of SMIM3 inhibited cell proliferation and colony formation in AML cells. The efficiency of SMIM3 knockdown in the Kasumi-1 and THP-1 cell lines was verified by A , B reverse transcription-quantitative polymerase chain reaction and E , F immune fluorescence, respectively. C , D Cell proliferation was detected by the CCK-8 assay in Kasumi-1 and THP-1. G , H The colony formation of SMIM3 -CTRL and SMIM3 -KD in Kasumi-1 and THP-1. CTRL, control; KD, knockdown; *P < 0.05 compared with CTRL cells; **P < 0.01 compared with CTRL cells; ***P < 0.001 compared with CTRL cells; Error bars indicate the standard deviation

Article Snippet: The human AML cell line Kasumi-1 was purchased from American Type Culture Collection (Manassas, VA, USA).

Techniques: Real-time Polymerase Chain Reaction, Fluorescence, CCK-8 Assay, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: Downregulation of SMIM3 inhibits growth of leukemia via PI3K-AKT signaling pathway and correlates with prognosis of adult acute myeloid leukemia with normal karyotype

doi: 10.1186/s12967-022-03831-8

Figure Lengend Snippet: Apoptosis and cell cycle analysis in AML cell lines. A , B The flow cytometry was applied to analyze apoptosis in SMIM3 knockdown cells compared with control. C – E Propidium iodide (PI) staining was applied to analyze the cell cycle of SMIM3 knockdown cells compared with control. F Alterations in apoptosis and cell cycle-related protein assay in SMIM3 knockdown cells compared with control. G , H Densitometric analysis of the WB signals. CTRL, control; KD, knockdown; *P < 0.05 compared with CTRL cells; **P < 0.01 compared with CTRL cells; ***P < 0.001 compared with CTRL cells

Article Snippet: The human AML cell line Kasumi-1 was purchased from American Type Culture Collection (Manassas, VA, USA).

Techniques: Cell Cycle Assay, Flow Cytometry, Staining

Journal: Journal of Translational Medicine

Article Title: Downregulation of SMIM3 inhibits growth of leukemia via PI3K-AKT signaling pathway and correlates with prognosis of adult acute myeloid leukemia with normal karyotype

doi: 10.1186/s12967-022-03831-8

Figure Lengend Snippet: GO/KEGG enrichment analysis of subjects with high or low SMIM3 expression in the TCGA-LAML dataset. A Volcano map of the DEGs, including 231up-regulated genes and 1889 down-regulated genes. B Heat map showing the top ten up-regulated and the top ten down-regulated genes. The samples are shown on the X-axis, and the DEGs are shown on the Y-axis. D Venn diagram of the overlap among the DEGs and correlated genes. C GO and KEGG enrichment analysis of the co-expressed genes of DEGs and correlated genes. MF, molecular function. CC, cellular component. BP, biological process. Different categories were shown on the Y-axis, and the X-axis reflected the percentage of DEGs. PI3K-AKT signaling pathway is critical for SMIM3 -mediated changes. G Western blot analysis of PI3K-AKT signaling pathway in SMIM3 -KD cells and controls. H , I Densitometric analysis of the WB signals. E SC79 (10 mM) reversed the inhibitory effect of SMIM3 knockdown on the proliferation of Kasumi-1, F at 24 h. CTRL, control; KD, knockdown; ***P < 0.001 compared with CTRL cells; Error bars indicate the standard deviation

Article Snippet: The human AML cell line Kasumi-1 was purchased from American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Western Blot, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: Downregulation of SMIM3 inhibits growth of leukemia via PI3K-AKT signaling pathway and correlates with prognosis of adult acute myeloid leukemia with normal karyotype

doi: 10.1186/s12967-022-03831-8

Figure Lengend Snippet: SMIM3 knockdown reduced the proliferation of AML cells in nude mice by inhibiting PI3K-AKT signaling pathway. A – D , F – I SMIM3 -KD inhibits tumor growth in Kasumi-1 and THP-1 cells. E , J The expression of PI3K-AKT signaling pathway proteins in tumor tissues by Western blot. CTRL, control; KD, knockdown; ***P < 0.001 compared with CTRL cells

Article Snippet: The human AML cell line Kasumi-1 was purchased from American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Western Blot