Journal: Oncotarget

Article Title: SALL1 expression in acute myeloid leukemia

doi: 10.18632/oncotarget.23448

Figure Lengend Snippet: SALL1 is expressed in human AML samples and cell lines ( A ) Positive nuclear IHC staining for SALL1 in AML compared to NBM. ( B ) RT-PCR and ( C ) qRT-PCR analysis of SALL1 expression in AML samples and AML cell lines as well as in NBM samples. The expression data were normalized to Rpl19 standard invariant gene levels. ( D ) Western blotting analyses of SALL1 protein expression in AML samples, and cell lines and in NBM sample. ( E ). Immunoprecipitation of RIPA buffer harvested total protein. Proteins were then analyzed by SDS-PAGE and membranes probed with monoclonal anti- SALL1 antibodies.

Article Snippet: Embryonic kidney cell line HEK293T, known to express SALL1 , and AML cell lines (MV4-11, THP-1,KG-1, K-562, TF-1, TF-1α) as well as CD34+ cells sorted from normal BM were purchased from ATCC ( American Type Culture Collection, Manassas, VA, USA).

Techniques: Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Western Blot, Immunoprecipitation, SDS Page

Journal: Oncotarget

Article Title: SALL1 expression in acute myeloid leukemia

doi: 10.18632/oncotarget.23448

Figure Lengend Snippet: Preferential SALL1 expression in LSCs enriched CD34+/CD38- cell subpopulation. ( A ) qRT-PCR analysis of SALL1 mRNA expression in sorted LSCs enriched CD34+/CD38- subpopulation compared to CD34+/CD38+ subpopulation ( B ). Western blotting analysis of SALL1 protein expression in sorted AML cells.

Article Snippet: Embryonic kidney cell line HEK293T, known to express SALL1 , and AML cell lines (MV4-11, THP-1,KG-1, K-562, TF-1, TF-1α) as well as CD34+ cells sorted from normal BM were purchased from ATCC ( American Type Culture Collection, Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Oncotarget

Article Title: SALL1 expression in acute myeloid leukemia

doi: 10.18632/oncotarget.23448

Figure Lengend Snippet: Exonal pattern of SALL1 expression on AML ( A ) Schematic illustration of SALL1 gene: All three exons of variant 1 are protein coding, while only exons 2 and 3 are in variant 2. ( B ). PCR of SALL1 genomic DNA and RT-PCR of SALL1 mRNA expression in HEK293, AML cells, AML cell lines and in NBM: All three exons of SALL1 are detected in genomic DNA of HEK293, AML cell lines, NBM, and AML. cDNA from SALL1 exon1 was expressed in HEK293 cells, but not in AML cells and cell lines. Exons 2 and 3 were detected in HEK293 cells and malignant cells. SALL1 mRNA was not detected in NBM. ( C ). Long-range RT-PCR using exon specific primers: exons 1, 2 and 3 were detected HEK293. Exons 2 and 3 but not exon1 were detected in AML TF1-α cell line at one length. SALL1 was not detected in NBM. Our primer is designed to detect the 3’ end of exon 2.

Article Snippet: Embryonic kidney cell line HEK293T, known to express SALL1 , and AML cell lines (MV4-11, THP-1,KG-1, K-562, TF-1, TF-1α) as well as CD34+ cells sorted from normal BM were purchased from ATCC ( American Type Culture Collection, Manassas, VA, USA).

Techniques: Expressing, Variant Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Oncotarget

Article Title: SALL1 expression in acute myeloid leukemia

doi: 10.18632/oncotarget.23448

Figure Lengend Snippet: ( A ) qRT-PCR and western blot analysis of SALL1 inhibition: HEK293 cells, 2 AML cell lines (MV4-11 and THP1) were stably infected and 1 AML primary patient sample was transiently infected with shRNAs of SALL1. Transfection efficiency was different amongst different cells ranging between >90% in MV4-11 to 40% in AML primary cells. ( B ) Cell proliferation: MV4-11 cells treated with sh-RNA against SALL1 or against GFP were incubated with cell proliferation WST-1 reagent. Sh- SALL1 treated cells showed a significant reduction in proliferation. And in ( C ) induced GM-colony formation. ( D ) In vivo engraftment of sh-SALL1 and of ctr-GFP MV4-11 and THP1 cells: SALL1 inhibition in MV4-11 cells resulted in AML engraftment and mice became morbid at an average of 5 weeks compared to mice injected with GFP-SALL1 MV4-11 cells which were alive and looked healthy at 6 months after treatment and with no evidence of leukemia when sacrificed. Spleen cells are sorted to HLA-DR and CD34 human AML markers. Less efficient SALL1 inhibition in THP1 cells resulted in delayed AML engraftment by an average of 9 weeks.

Article Snippet: Embryonic kidney cell line HEK293T, known to express SALL1 , and AML cell lines (MV4-11, THP-1,KG-1, K-562, TF-1, TF-1α) as well as CD34+ cells sorted from normal BM were purchased from ATCC ( American Type Culture Collection, Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Western Blot, Inhibition, Stable Transfection, Infection, Transfection, Incubation, In Vivo, Injection

Journal: Oncotarget

Article Title: SALL1 expression in acute myeloid leukemia

doi: 10.18632/oncotarget.23448

Figure Lengend Snippet: SALL1 inhibition resulted in upregulation of PTEN expression and inhibition of down-stream m-TOR and in downregulation of B-catenin and NF-κB: ( A, B ) SALL1 inhibition resulted in upregulation of PTEN and inhibition of m-TOR in MV4-11 and primary AML, more prominent with MV4-11 where SALL1 downregulation efficiency was almost 100%, than in primary AML. ( C ) In genomic PTEN deleted THP1 cells, mTOR did not change. ( D ) SALL1 inhibition is shown with decreased B-catinin, and NF-κB expression in in both MV4-11 and THP1.

Article Snippet: Embryonic kidney cell line HEK293T, known to express SALL1 , and AML cell lines (MV4-11, THP-1,KG-1, K-562, TF-1, TF-1α) as well as CD34+ cells sorted from normal BM were purchased from ATCC ( American Type Culture Collection, Manassas, VA, USA).

Techniques: Inhibition, Expressing