Journal: Nature Communications

Article Title: THRAP3 promotes ferroptosis resistance in acute myelocytic leukemia through SLU7-mediated alternative splicing of GIT2

doi: 10.1038/s41467-025-66931-1

Figure Lengend Snippet: A Up-regulation of THRAP3 was identified in relapsed AML patients ( n = 416) and primary AML patients ( n = 2128) from TARGET database. The box plots show the median (centre line), the 25th and 75th percentiles (box limits), and the minima and maxima. B TCGA database analyzed the correlation between THRAP3 expression and survival of AML patients ( n = 132). C RT-qPCR analysis of THRAP3 expression in bone marrow (BM) of healthy volunteers and AML patients (normal group, n = 12; AML group, n = 82). D THRAP3 expression in AML patients from newly diagnosed ( n = 82), complete remission (CR) ( n = 42) and relapse groups ( n = 16) was assessed by RT-qPCR. E Kaplan-Meier evaluated the correlation between THRAP3 expression and overall survival of AML patients (THRAP3 high expression, n = 41; THRAP3 low expression, n = 41). F Western blotting determined THRAP3 protein level in normal and AML samples ( n = 12). N, normal BM samples; A, BM samples of AML patients. G , H RT-qPCR and Western blotting analysis of THRAP3 expression in normal BMSCs, BMMNCs, HS-5 and multiple AML cell lines ( n = 3, biological replicates). The data are represented as mean ± standard deviation (SD). Experiments were repeated independently for 3 times with similar results, and representative image from one experiment is shown in ( F , H ). One-way ANOVA followed by Tukey’s post hoc test (for D , G , H ), two-tailed Student’s t test (for A , C , F ) and Log-rank test (for B , E ) were performed for statistical analysis. * p < 0.05, ** p < 0.01, *** p < 0.001. Source data are provided as a Source Data file.

Article Snippet: AML cell line NB4 (Cat. No.: CL-0676) was purchased from Procell (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Two Tailed Test

Journal: Nature Communications

Article Title: THRAP3 promotes ferroptosis resistance in acute myelocytic leukemia through SLU7-mediated alternative splicing of GIT2

doi: 10.1038/s41467-025-66931-1

Figure Lengend Snippet: THP-1 cells were transfected with sgTHRAP3 or sgNC; HL-60 cells were transfected with THRAP3 expression plasmid or vector. A CCK-8 evaluated AML cell proliferation. Orthotopic AML xenografts were generated by injection with HL-60 cells into tail veins of NOD/SCID mice. B Survival of mice was monitored. C , D livers and spleens were collected from mice and weighed. E The percentage of CD45 + Ki67 + in livers and spleens was detected by flow cytometry. n = 3, biological replicates for A; n = 6 mice/group for ( B – E ). The data are represented as mean ± standard deviation (SD). Experiments were repeated independently for 6 times with similar results, and representative images from one experiment are shown in ( E ). Two-tailed Student’s t test (for A , D , E ) and Log-rank test (for B ) was performed for statistical analysis. ** p < 0.01, *** p < 0.001. Source data are provided as a Source Data file.

Article Snippet: AML cell line NB4 (Cat. No.: CL-0676) was purchased from Procell (Wuhan, China).

Techniques: Transfection, Expressing, Plasmid Preparation, CCK-8 Assay, Generated, Injection, Flow Cytometry, Standard Deviation, Two Tailed Test

Journal: Open Medicine

Article Title: High STAT4 expression correlates with poor prognosis in acute myeloid leukemia and facilitates disease progression by upregulating VEGFA expression

doi: 10.1515/med-2023-0840

Figure Lengend Snippet: SiSTAT4 decreased STAT4 expression, while overexpressed STAT4 did conversely in transfected HL60 and NB4 cells. (a) The expression of STAT4 in HL60 cells after transfection was examined by qPCR, and GAPDH was used as the loading gene. (b) The expression of STAT4 in HL60 cells after transfection was examined by Western blot, and GAPDH was adopted as the internal control. (c) The expression of STAT4 in NB4 cells after transfection was examined by qPCR, and GAPDH was employed as a loading gene. (d) The expression of STAT4 in NB4 cells after transfection was examined by Western blot, and GAPDH was utilized as the internal control. (e) The expression of STAT4 in HL60 cells after transfection was examined by qPCR, and GAPDH was harnessed as the reference gene. (f) The expression of STAT4 in HL60 cells after transfection was measured by Western blot, and GAPDH was exploited as the internal control. (g) The expression of STAT4 in NB4 cells after transfection was quantified by qPCR, and GAPDH was applied as the reference gene. (h) The expression of STAT4 in NB4 cells after transfection was determined by Western blot, and GAPDH was adopted as the loading control. *** P < 0.001 vs siNC; ^^^ P < 0.001 vs NC (NC: negative control, siSTAT4: siRNA targeting STAT4, siNC: negative control of siSTAT4).

Article Snippet: Human AML cell lines including HL60 (CL-0110), KG1 (CL-0132), Kasumi-1 (CL-0556), and NB4 (CL-0676) were purchased from Procell (Wuhan, China), and normal bone marrow cell line HS-5 (CRL-11882) was ordered from American Type Culture Collection (ATCC; Maryland, USA).

Techniques: Expressing, Transfection, Western Blot, Control, Negative Control

Journal: Open Medicine

Article Title: High STAT4 expression correlates with poor prognosis in acute myeloid leukemia and facilitates disease progression by upregulating VEGFA expression

doi: 10.1515/med-2023-0840

Figure Lengend Snippet: STAT4 silencing decreased the viability, while overexpressed STAT4 did conversely in transfected AML cells. (a–d) The viability of HL60 and NB4 cells after transfection was examined by MTT assay. * P < 0.05, *** P < 0.001 vs siNC; ^ P < 0.05, ^^^ P < 0.001 vs NC.

Article Snippet: Human AML cell lines including HL60 (CL-0110), KG1 (CL-0132), Kasumi-1 (CL-0556), and NB4 (CL-0676) were purchased from Procell (Wuhan, China), and normal bone marrow cell line HS-5 (CRL-11882) was ordered from American Type Culture Collection (ATCC; Maryland, USA).

Techniques: Transfection, MTT Assay

Journal: Open Medicine

Article Title: High STAT4 expression correlates with poor prognosis in acute myeloid leukemia and facilitates disease progression by upregulating VEGFA expression

doi: 10.1515/med-2023-0840

Figure Lengend Snippet: STAT4 silencing decreased Bcl-2 level, yet increased apoptosis and Bax level in AML cells, while overexpressed STAT4 did conversely. (a and b) The apoptosis of HL60 and NB4 cells after transfection was examined by flow cytometry. (c) The expressions of Bcl-2 and Bax in HL60 cells after transfection were quantified by qPCR, and GAPDH was used as the reference gene. (d) The expressions of Bcl-2 and Bax in HL60 cells after transfection were detected by Western blot, and GAPDH was used as the loading control. (e) The expressions of Bcl-2 and Bax in NB4 cells after transfection were measured by qPCR, and GAPDH was used as the reference gene. (f) The expressions of Bcl-2 and Bax in NB4 cells after transfection were examined by Western blot, and GAPDH was used as the internal control. *** P < 0.001 vs siNC; ^^^ P < P < 0.001 vs NC.

Article Snippet: Human AML cell lines including HL60 (CL-0110), KG1 (CL-0132), Kasumi-1 (CL-0556), and NB4 (CL-0676) were purchased from Procell (Wuhan, China), and normal bone marrow cell line HS-5 (CRL-11882) was ordered from American Type Culture Collection (ATCC; Maryland, USA).

Techniques: Transfection, Flow Cytometry, Western Blot, Control

Journal: Open Medicine

Article Title: High STAT4 expression correlates with poor prognosis in acute myeloid leukemia and facilitates disease progression by upregulating VEGFA expression

doi: 10.1515/med-2023-0840

Figure Lengend Snippet: STAT4 silencing decreased the angiogenesis and VEGFA level, while overexpressed STAT4 did conversely. (a and b) The angiogenesis of HL60 and NB4 cells after transfection was examined by tube formation assay under ×100 magnification. (c) The expressions of STAT4 and VEGFA in HL60 cells after transfection were tested by qPCR, and GAPDH was used as the reference gene. (d) The expressions of STAT4 and VEGFA in HL60 cells after transfection were determined by Western blot, and GAPDH was adopted as the internal control. (e) The expressions of STAT4 and VEGFA in NB4 cells after transfection were assayed by qPCR, and GAPDH was utilized as the reference gene. (f) The expressions of STAT4 and VEGFA in NB4 cells after transfection were measured by Western blot, and GAPDH was exploited as the loading control. *** P < 0.001 vs siNC; ^^^ P < 0.001 vs NC (VEGFA: vascular endothelial growth factor A).

Article Snippet: Human AML cell lines including HL60 (CL-0110), KG1 (CL-0132), Kasumi-1 (CL-0556), and NB4 (CL-0676) were purchased from Procell (Wuhan, China), and normal bone marrow cell line HS-5 (CRL-11882) was ordered from American Type Culture Collection (ATCC; Maryland, USA).

Techniques: Transfection, Tube Formation Assay, Western Blot, Control

Journal: Open Medicine

Article Title: High STAT4 expression correlates with poor prognosis in acute myeloid leukemia and facilitates disease progression by upregulating VEGFA expression

doi: 10.1515/med-2023-0840

Figure Lengend Snippet: VEGFA silencing counteracted the effects of overexpressed STAT4 on promoting the viability and angiogenesis as well as inhibiting the apoptosis of AML cells. (a and b) The viability of HL60 and NB4 cells after transfection was examined by MTT assay. (c and d) The angiogenesis of HL60 and NB4 cells after transfection was determined by tube formation assay under ×100 magnification. (e and f) The apoptosis of HL60 and NB4 cells after transfection was tested by flow cytometry. * P < 0.05, ** P < 0.01 , *** P < 0.001 vs NC + vector; ^ P < 0.05, ^^ P < 0.01, ^^^ P < 0.001 vs STAT4 + vector; # P < 0.05, ## P < 0.01, ### P < 0.001 vs NC + siVEGFA (siVEGFA: siRNA targeting VEGFA, vector: negative control for siVEGFA).

Article Snippet: Human AML cell lines including HL60 (CL-0110), KG1 (CL-0132), Kasumi-1 (CL-0556), and NB4 (CL-0676) were purchased from Procell (Wuhan, China), and normal bone marrow cell line HS-5 (CRL-11882) was ordered from American Type Culture Collection (ATCC; Maryland, USA).

Techniques: Transfection, MTT Assay, Tube Formation Assay, Flow Cytometry, Plasmid Preparation, Negative Control