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( A ) Analysis of <t>KLF4</t> gene methylation from the Cancer Cell Line Encyclopedia (CCLE) in a panel of hematologic malignancy cell lines. ( B ) Correlation of KLF4 transcript levels and DNA methylation in AML and non-AML cell lines. Linear regression analysis was conducted with 95% confidence interval.

( A ) Analysis of <t>KLF4</t> gene methylation from the Cancer Cell Line Encyclopedia (CCLE) in a panel of hematologic malignancy cell lines. ( B ) Correlation of KLF4 transcript levels and DNA methylation in AML and non-AML cell lines. Linear regression analysis was conducted with 95% confidence interval.

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1) Product Images from "Krüppel-like factor 4 promotes survival and expansion in acute myeloid leukemia cells"

Article Title: Krüppel-like factor 4 promotes survival and expansion in acute myeloid leukemia cells

Journal: Oncotarget

doi: 10.18632/oncotarget.27878

( A ) Analysis of KLF4 gene methylation from the Cancer Cell Line Encyclopedia (CCLE) in a panel of hematologic malignancy cell lines. ( B ) Correlation of KLF4 transcript levels and DNA methylation in AML and non-AML cell lines. Linear regression analysis was conducted with 95% confidence interval.

Figure Legend Snippet: ( A ) Analysis of KLF4 gene methylation from the Cancer Cell Line Encyclopedia (CCLE) in a panel of hematologic malignancy cell lines. ( B ) Correlation of KLF4 transcript levels and DNA methylation in AML and non-AML cell lines. Linear regression analysis was conducted with 95% confidence interval.

Techniques Used: Methylation, DNA Methylation Assay

( A ) Relative KLF4 expression was measured by qPCR in AML, CML, and LCL cell lines cultured in the presence of 250 and 500 nM 5-Aza for 96 hours to induce DNA demethylation. Data represents relative mRNA expression (DDC T ) expressed as mean ± s.d. ( n = 3). ( B ) Immunoblot analysis of KLF4 expression after treatment with 5-Aza for 96 hours. Actin was used as a loading control. ( C ) Immunoblot analysis of KLF4 and MYC expression in cell lines treated with 10 μM of MG-132 for 4 hours. Actin and GAPDH were used as loading control. Data shown are representative of two independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, two-tailed Student’s t -test.

Figure Legend Snippet: ( A ) Relative KLF4 expression was measured by qPCR in AML, CML, and LCL cell lines cultured in the presence of 250 and 500 nM 5-Aza for 96 hours to induce DNA demethylation. Data represents relative mRNA expression (DDC T ) expressed as mean ± s.d. ( n = 3). ( B ) Immunoblot analysis of KLF4 expression after treatment with 5-Aza for 96 hours. Actin was used as a loading control. ( C ) Immunoblot analysis of KLF4 and MYC expression in cell lines treated with 10 μM of MG-132 for 4 hours. Actin and GAPDH were used as loading control. Data shown are representative of two independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, two-tailed Student’s t -test.

Techniques Used: Expressing, Cell Culture, Western Blot, Control, Two Tailed Test

( A ) Multiplex sgRNA targeting strategy to induce KLF4 gene deletion via CRISPR/CAS9 technology and comparison of sequencing tracks of parental and edited leukemic cells. ( B ) Cell growth of NB4 KO and MM6 KO cells compared to parental cell lines ( n = 3); inset shows loss of KLF4 expression in edited cell lines. ( C ) Flow cytometric detection of Annexin V and 7AAD in NB4 and NB4 KO cells. Representative plots are included on the left and statistical analysis on the right ( n = 3). ( D ) Cell cycle analysis of NB4 and NB4 KO by flow cytometric detection of propidium iodine staining of nuclei ( n = 3). ( E and F ) Apoptosis and cell cycle analysis for MM6 and MM6 KO cells. Data represent mean ± s.d. ( n = 3). * p < 0.05, *** p < 0.001, two-tailed Student’s t -test.

Figure Legend Snippet: ( A ) Multiplex sgRNA targeting strategy to induce KLF4 gene deletion via CRISPR/CAS9 technology and comparison of sequencing tracks of parental and edited leukemic cells. ( B ) Cell growth of NB4 KO and MM6 KO cells compared to parental cell lines ( n = 3); inset shows loss of KLF4 expression in edited cell lines. ( C ) Flow cytometric detection of Annexin V and 7AAD in NB4 and NB4 KO cells. Representative plots are included on the left and statistical analysis on the right ( n = 3). ( D ) Cell cycle analysis of NB4 and NB4 KO by flow cytometric detection of propidium iodine staining of nuclei ( n = 3). ( E and F ) Apoptosis and cell cycle analysis for MM6 and MM6 KO cells. Data represent mean ± s.d. ( n = 3). * p < 0.05, *** p < 0.001, two-tailed Student’s t -test.

Techniques Used: Multiplex Assay, CRISPR, Comparison, Sequencing, Expressing, Cell Cycle Assay, Staining, Two Tailed Test

Cytotoxicity was determined using Cell-Titer Glo cytotoxicity assay after 48 h treatment with either daunorubicin or ara-C. ( A ) Cytotoxicity of NB4 and NB4 KO cells to daunorubicin ( n = 3). ( B ) Cytotoxicity of NB4 and NB4 KO cells to ara-C ( n = 3). ( C ) Cytotoxicity of MM6 and MM6 KO cells to daunorubicin ( n = 3). ( D ) Cytotoxicity of MM6 and MM6 KO cells to ara-C ( n = 3). Expression of KLF4 protein in parental cells in response to treatment at IC 50 dose is shown in figure insets. Data represent mean ± s.d.

Figure Legend Snippet: Cytotoxicity was determined using Cell-Titer Glo cytotoxicity assay after 48 h treatment with either daunorubicin or ara-C. ( A ) Cytotoxicity of NB4 and NB4 KO cells to daunorubicin ( n = 3). ( B ) Cytotoxicity of NB4 and NB4 KO cells to ara-C ( n = 3). ( C ) Cytotoxicity of MM6 and MM6 KO cells to daunorubicin ( n = 3). ( D ) Cytotoxicity of MM6 and MM6 KO cells to ara-C ( n = 3). Expression of KLF4 protein in parental cells in response to treatment at IC 50 dose is shown in figure insets. Data represent mean ± s.d.

Techniques Used: Cytotoxicity Assay, Expressing

Figure Legend Snippet:

Techniques Used:

Expressing:

Article Title: Cytomegalovirus infection of the fetal brain: intake of aspirin during pregnancy blunts neurodevelopmental pathogenesis in the offspring
Article Snippet: .. There is more prominent expression of Ptgs1 than Ptgs2 in mouse microglia, according to RNAseq databases ( https://www.brainrnaseq.org/ ; [ ] ( http://neuroexpresso.org/ ,[ ]), and notably during brain development, as shown by our in silico search at the Broad Institute website ( https://singlecell.broadinstitute.org/single_cell/study/SCP1290 , [ ]. ..

Article Title: Krüppel-like factor 4 promotes survival and expansion in acute myeloid leukemia cells
Article Snippet: .. KLF4 mRNA expression (RNASeq) and DNA methylation (RRBS) were downloaded for cell lines from the publicly accessible cancer cell line encyclopedia database from Broad Institute. .. Data were replotted and formatted using GraphPad prism software, and linear regression with coefficient of correlation was calculated.

RNA sequencing:

In Silico:

Sequencing:

Article Title: Upregulated heme biosynthesis increases obstructive sleep apnea severity: a pathway-based mendelian randomization study
Article Snippet: .. Genome sequencing and RNASeq for “NHLBI TOPMed: MESA and MESA Family AA-CAC” (phs001416.v2.p1) was performed at the Broad Institute Genomics Center (3U54HG003067-13S1, HHSN268201600034I). ..

DNA Methylation Assay:

other:

Article Title: Allele-specific silencing of the gain-of-function mutation in Huntington’s disease using CRISPR/Cas9
Article Snippet: Then, genome-wide RNAseq analysis was performed by the Broad Institute.

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