Journal: Cell Death & Disease

Article Title: The long noncoding RNA HOTAIRM1 controlled by AML1 enhances glucocorticoid resistance by activating RHOA/ROCK1 pathway through suppressing ARHGAP18

doi: 10.1038/s41419-021-03982-4

Figure Lengend Snippet: A Heatmap of the genes expression level, which may be regulated by HOTAIRM1 and contribute to GC resistance. B ARHGAP18 were selected by overlapping the ChIRP target genes and the genes, which became similar to K-S after HOTAIM1 knockdown in K-R. C The expression levels of ARHGAP18 were measured in K-S-scramble, K-R-scramble, and K-R with HOTAIRM1 knockdown using qRT-PCR (left). HOTAIRM1 was not affected by the knockdown of ARHGAP18 in K-S (right). D Tracks of the HOTAIRM1 binding peaks near the ARHGAP18 gene locus in ChIRP-seq data. The dark blue represents the even probe group, the purple represents the odd probe group, the red represents the peak of the merged odd and even probe data, the gray represents the LacZ probe group, and the black represents the input, the red dashed box represents the binding peak. E K-R cells were transfected with the HOTAIRM1 binding sequence of ARHGAP18 in luciferase reporter assay. Luciferase activity was compared with that of the empty construct with pGL3 promoter. F pGL3-ARHGAP18 were co-transfected into K-R cells with either nontargeting control vectors or HOTAIRM1-targeting shRNA vectors. Data information: Data were represented as mean ± SD, n = 3 ( C, E–F ). Difference between groups was assessed using Student’s t -test: * p < 0.05, ** p < 0.01, *** p < 0.001, ns means no statistical significance.

Article Snippet: Lentiviral vectors expressing short hairpin RNAs (shRNAs) targeting the selected HOTAIRM1 sites (Supplementary Table ) were constructed by inserting each shRNA cassette into a pLKO-TRC backbone plasmid (Addgene plasmid 10878).

Techniques: Expressing, Knockdown, Quantitative RT-PCR, Binding Assay, Transfection, Sequencing, Luciferase, Reporter Assay, Activity Assay, Construct, Control, shRNA

Journal: PLoS ONE

Article Title: MSH3-Deficiency Initiates EMAST without Oncogenic Transformation of Human Colon Epithelial Cells

doi: 10.1371/journal.pone.0050541

Figure Lengend Snippet: (A) Western blot analysis of HCT116, HCT116+chr3 and HCEC-1CT cells. (B) Silencing of MMR-protein MSH3 by gene-specific shRNA in HCEC-1CT-[AAAG]17 cells resulted in 70% repression of MSH3. MSH6 was not affected.

Article Snippet: HCEC-1CT, HCEC-1CT-[AAAG]17 and HCEC-1CT-[CA]13 cells were transfected with the pRFP-C-RS vectors (Origene) either without shRNA cassette insert (control), a non-effective 29-mer scrambled (scrambled) or a gene-specific shRNA cassette for suppression of MSH3 (shMSH3).

Techniques: Western Blot, shRNA