kansl2  (Proteintech)

Journal: Cell reports

Article Title: Comprehensive synergy mapping links a BAF- and NSL-containing “supercomplex” to the transcriptional silencing of HIV-1

doi: 10.1016/j.celrep.2023.113055

Figure Lengend Snippet: (A) Design of a two-vector system for introducing a pair of sgRNAs into each cell. (B) A schematic of REACTS to screen the genome for sgRNAs synergizing with eight query sgRNAs targeting known host repressors of latent HIV. (C) Example primary FACS data of synergy between sgNFKBIA and sgBCL7C. (D) An overview of the Bliss independence test used to call synergy. (E) REACTS identified 32 synergistic, 6 additive, and 2 antagonistic pairs of sgRNAs. Red, pairs involving BCL7C, KANSL2, and SIRT2. (F) A network representing the individually validated synergies identified by REACTS. Related to and and and .

Article Snippet: siRNA targeting human KANSL2 , Thermo Fisher , s29791.

Techniques: Plasmid Preparation

Journal: Cell reports

Article Title: Comprehensive synergy mapping links a BAF- and NSL-containing “supercomplex” to the transcriptional silencing of HIV-1

doi: 10.1016/j.celrep.2023.113055

Figure Lengend Snippet: (A–R) J-Lat A2 cells were nucleofected with siRNAs targeting the indicated genes or plasmid vectors expressing the indicated proteins and subjected to ChIP-qPCR using normal rabbit immunoglobulin G (IgG) (for control) or antibodies that specifically recognize the indicated proteins to quantify their occupancy of the Nuc-1 region of HIV LTR. The ChIP-qPCR signals were normalized to input DNA. Error bars represent mean ± SD from three experimental replicates. *p < 0.05, **p < 0.01, and ***p < 0.001 by two-tailed Student’s t test. For (B)–(P), significance tests were conducted between each test group versus the control groups (siNegative or empty vector). For experiments in (B)–(H) and (Q), aliquots of the nucleofected cells were subjected to western blot analyses (A and R, respectively) to determine levels of the indicated proteins. (S) Models showing the different mechanisms by which BCL7C, KANSL2, and SIRT2 reduce HIV transcription. Related to .

Article Snippet: siRNA targeting human KANSL2 , Thermo Fisher , s29791.

Techniques: Plasmid Preparation, Expressing, ChIP-qPCR, Control, Two Tailed Test, Western Blot

Journal: Cell reports

Article Title: Comprehensive synergy mapping links a BAF- and NSL-containing “supercomplex” to the transcriptional silencing of HIV-1

doi: 10.1016/j.celrep.2023.113055

Figure Lengend Snippet:

Article Snippet: siRNA targeting human KANSL2 , Thermo Fisher , s29791.

Techniques: Control, Virus, Recombinant, Protease Inhibitor, Negative Control, Reverse Transcription, SYBR Green Assay, Genome Wide, Software

Journal: Nature Communications

Article Title: Evolutionary conserved NSL complex/BRD4 axis controls transcription activation via histone acetylation

doi: 10.1038/s41467-020-16103-0

Figure Lengend Snippet: a RT-qPCR analyses of Kansl2 fl/fl , Cre-ERT2 mESCs with ( Kansl2 KO) and without ( Kansl2 WT) tamoxifen treatment (500 nM, 3 days), and dBET6-treated (100 nM, 4 h) Kansl2 WT cells, see also Supplementary Fig. . Expression is normalized to HPRT and relative to Kansl2 WT. b ChIP-qPCR of BRD4 in Kansl2 KO, Kansl2 WT and dBET6-treated mESCs. ChIP enrichments are relative to gene desert signal. Primers target KANSL2-responsive (target) and KANSL2-non-responsive (control) genes. a , b Bars represent mean ± SEM ( n = 3, for dBET ChIP n = 2 biological replicates). Source data are provided as a Source Data file. c Heatmap of row-scaled normalized RNA-seq counts from fibroblast cell lines of three Koolen-de Vries/ KANSL1 haploinsufficient ( KANSL1 +/−) patients and four controls (for genes FDR < 0.2). Order was generated by unsupervised hierarchical clustering. Genes significantly downregulated (DE down) (FDR < 0.05) upon JQ1 treatment in MOLT4 cells are indicated in black, upregulated (DE up) or not differentially expressed (not DE) in white. d Left, log2 fold changes of conserved, NSL complex-dependent genes for NSL1 RNAi in Drosophila (FDR < 0.05), and Kansl3 knockdown in mESCs (FDR < 0.05, ). Right, row-scaled normalized RNA-seq counts of KANSL1 +/− patients and controls (FDR < 0.4). Red dot indicates association with GO term Metabolic Process. e Ingenuity Pathway Analysis showing most affected Disease and Functions groups of DE genes (FDR < 0.1) obtained from DESeq2 analysis of KANSL1 +/− patient fibroblast RNA-seq. Grey Z-score indicates NA. Boxes are sized by negative log p- value. f Boxplots of log2 fold change of KANSL1 +/− patient fibroblasts versus controls for classes of genes, based on differential expression upon JQ1 treatment : DE down ( n = 5581), DE up ( n = 1482), not DE ( n = 3716). Boxplots show median (centre), interquartile-range (box) and minima/maxima (whiskers).Two-sided Wilcoxon-rank-sum test was applied. g Left and middle: percentage of genes whose mouse orthologues are promoter-bound by KANSL3 ( n = 3665)(left) or MSL2 ( n = 846) (middle) in mESC and downregulated (black bars) or upregulated (grey bars) in KANSL1 +/− patients. Right: percentage of genes significantly downregulated (FDR < 0.05) upon JQ1 treatment in MOLT4 cells ( n = 5581) and down-or upregulated in KANSL1 +/− patients. Up/down classification of patient gene expression indicates directionality. Overrepresentation of downregulated genes was tested with one-sided Fisher’s exact test.

Article Snippet: Following primary antibodies were used: dBRD4 (Paro lab, ID166, 1:1000), Pol2 ser2p (ab5095, Abcam, 1:5000), Pol2 ser5p (ab5131, Abcam, 1:5000), Rpb3 (Akhtar lab, 1:1000), H3 (ab10799, Abcam, 1:3000), NSL3 (Akhtar lab, 1:1000), MOF (Akhtar lab, 1:3000), H2A.V (61752, Active Motif, 1:2000), MCRS2 (Akhtar lab, 1:3000), H4K16ac (07-329, Millipore, 1:3000), KANSL2 (HPA038497, Sigma, 1:1000), BRD4(A301-985, Bethyl, 1:2000), ACTIN-HRP(sc-1616, Santa Cruz, 1:5000), GAPDH-HRP (MA5-15738, Thermo Scientific, 1:5000), FLAG-HRP(A8592, Sigma, 1:5000).

Techniques: Quantitative RT-PCR, Expressing, ChIP-qPCR, Control, RNA Sequencing, Generated, Knockdown, Quantitative Proteomics, Gene Expression

Journal: Scientific Reports

Article Title: KANSL2 and MBNL3 are regulators of pancreatic ductal adenocarcinoma invasion

doi: 10.1038/s41598-020-58448-y

Figure Lengend Snippet: MBNL3 and KANSL2 regulate PANC-1 invasion. ( A ) The downregulation of genes giving the strongest response from the hit validation for invasion, MBNL3 and KANSL2 , were confirmed by shRNA-mediated knockdown in response to doxycycline versus no treatment. ( B ) Images are representative of the observed phenotype, and the yellow pseudo-colored cells are those that have invaded the lower chamber of the invasion Boyden Chamber. ( C ) The graphs represent the knockdown efficiency of KANSL2 and MBNL3 as assessed by the mRNA levels. The data presented are based on three independent experiments, and the P -values were determined using ANOVA with Tukey’s HSD ** P < 0.01, **** P < 0.0001.

Article Snippet: 18S and KANSL2 and MBNL3 TaqMan probes were purchased from Thermo Fisher Scientific (Waltham, MA).

Techniques: shRNA