Journal: Oxidative Medicine and Cellular Longevity

Article Title: METTL14 Regulates Intestine Cellular Senescence through m 6 A Modification of Lamin B Receptor

doi: 10.1155/2022/9096436

Figure Lengend Snippet: LBR is a key downstream gene of METTL14. (a) Schematic diagram showing the screening criterion for METTL14 targets. Venn diagram of all the possible overlapping transcripts in nine comparison groups sequenced together, showing reduced abundance with senescence and filtering for a significant FDR < 0.15. RNA-seq data was acquired from GEO datasets (GSE130727). RT-qPCR was performed in CCD18-Co with METTL14 knockdown to validate the overlapped genes. The LBR was consistent among 19 genes. FBL : fibrillarin; ITPRIP : inositol 1,4,5-trisphosphate receptor interacting protein; LBR : lamin B receptor; FAM129A : family with sequence similarity 129 member A; MCUB: mitochondrial calcium uniporter dominant negative subunit beta; SSRP1: structure specific recognition protein 1; TMSB15A : thymosin beta 15A; HIST1H1D : H1.3 linker histone, cluster member; CDCA7L: cell division cycle associated 7 like; CBS : cystathionine beta-synthase; HIST1H1A: H1.1 linker histone, cluster member; HIST1H1E : H1.4 linker histone, cluster member; PTMA : prothymosin alpha; ANP32B : acidic nuclear phosphoprotein 32 family member B; CBX2 : chromobox 2; HIST2H2AB: histone cluster 2 H2A family member B; PARP1 : poly(ADP-ribose) polymerase 1; H3FF : H3 clustered histone 11; H2BH : H2B clustered histone 9. (b) The expression of LBR following METTL14 knockdown or overexpression was evaluated by western blotting. (c) Prediction of the specific m 6 A methylation loci of LBR using the SRAMP website. (d) METTL14-silenced cells were treated with actinomycin D and harvested at 0, 2, or 4 h. RNA decay rates were determined to estimate the stability of LBR mRNA (normalized to the expression at 0 h). Data are expressed as mean ± SD of three independent biological experiments. Two-way ANOVA analysis. ∗∗ p < 0.01. (e) m 6 A modification of LBR was detected by MeRIP-qPCR analysis using anti-IgG and anti-m 6 A antibodies. Relative m 6 A enrichment of LBR mRNA for each IP group was normalized to input. Data are expressed as mean ± SD of three independent biological experiments. One-way ANOVA followed by Tukey's test. ∗∗ p < 0.01.

Article Snippet: After blocking with 5% BSA solutions for 2 h at room temperature, membranes were incubated using the following antibodies: LBR (diluted 1 : 500, ab32535; Abcam, Cambridge, UK), p53 (diluted 1 : 1000, ab26; Abcam), p21 (diluted 1 : 1000, ab109199; Abcam), p16 (diluted 1 : 1000, ET1602-9; HUABIO, Hangzhou, China), METTL14 (diluted 1 : 1000, 26158-1-AP; Proteintech, Wuhan, China), WTAP (diluted 1 : 1000, 60188-1-Ig; Proteintech), ALKBH5 (diluted 1 : 1000, 16837-1-AP; Proteintech), YTHDC1 (diluted 1 : 1000, A7318; ABclonal, Wuhan, China), YTHDC2 (diluted 1 : 1000, A15004; ABclonal), METTL3 (diluted 1 : 1000, db318; Diagbio, Hangzhou, China), and β -actin (as the loading control) (diluted 1 : 1000, 3700; CST, Danvers, USA) at 4°C overnight.

Techniques: RNA Sequencing Assay, Quantitative RT-PCR, Sequencing, Dominant Negative Mutation, Expressing, Over Expression, Western Blot, Methylation, Modification

Journal: AMIA Annual Symposium Proceedings

Article Title: Patient-Specific Models for Predicting the Outcomes of Patients with Community Acquired Pneumonia

doi:

Figure Lengend Snippet: Performance of the various methods on the test set of 686 cases. The method for computing the errors in column 3 is described in detail in the text. The last column gives the number of patients that would be treated at home out of 686 when operating at the point on the ROC curve that corresponds to the error rate in the third column. The last row gives the actual error rate and the actual number of patients treated at home.

Article Snippet: We implemented the modified LBR in Matlab (version 7) and the PSA in Java.

Techniques: Modification

Journal: AMIA Annual Symposium Proceedings

Article Title: Patient-Specific Models for Predicting the Outcomes of Patients with Community Acquired Pneumonia

doi:

Figure Lengend Snippet: ROC curve on the test set for the modified LBR method. The dot indicates the operating point on the curve that is discussed in the text.

Article Snippet: We implemented the modified LBR in Matlab (version 7) and the PSA in Java.

Techniques: Modification