Fig. 1: MIR31HG knock-down decreases the induction of SASP components during BRAF-induced senescence.
From: The long non-coding RNA MIR31HG regulates the senescence associated secretory phenotype
a Relative MIR31HG expression normalized to housekeeping genes (HPRT1 and RPLP0) in BJ ER:BRAF cells were treated with ethanol (Control) or 1 μM 4-OHT for 48 h (n = 4). b Box plot showing MIR31HG expression in log2 (fpkm + 1) units in thyroid carcinoma (THCA) and colorectal carcinoma (CRC) comparing BRAF mutant and BRAF wild-type tumours. The box plot represents the median (middle line), the box indicates the first and third quartiles and the whiskers indicate ±1.5 × interquartile range. Two-tailed Wilcoxon test was performed to compare the expression differences in the BRAF mutant and BRAF wild-type tumours (CRC BRAF_wt = 309; CRC BRAF_mutant = 49; THCA BRAF_mutant = 290; THCA BRAF_wt = 199). c Heat map showing relative expression of differentially expressed genes in BJ ER:BRAF cells (control or siMIR31HG), treated with ethanol (Control 1–3) or 1 μM 4-OHT for 48 h (Sen 1–3 and Sen siMIR31HG 1–3). d Heat map showing relative expression in RPKMs of a subset of SASP genes previously defined13 from the data provide in a, where differentially expressed genes (‘DE’, FDR < 0.01) are indicated in red, unchanged (non-significant, ‘ns’) in white. e qRT-PCR analysis of selected components of the SASP normalized to housekeeping genes (HPRT1 and RPLP0) in BJ ER:BRAF cells transfected with the indicated siRNAs (Control or siMIR31HG1-2), treated with ethanol (Control) or 1 μM 4-OHT for 48 h. The graphs show results compared to control ethanol-treated set to 1 (n = 4). f Mass spectrometry-based secretome analysis of senescent BJ ER:BRAF (1 μM 4-OHT) treated with siMIR31HG or control siRNA. The volcano plot shows differentially secreted proteins in MIR31HG knock-down senescent cells (Sen siMIR31HG) compared to control senescence cells (Sen) (−log10 p value along y-axis and log fold change along x-axis). Names are displayed for significantly changed proteins between secretomes (pval < 0.01) and SASP proteins are marked in red. g Secreted IL6, CXCL1, IL8 and MMP3 (pg/ml) measured by ELISA in the CM of BJ ER:BRAF (Control or siMIR31HG1-2) treated for 72 h with ethanol (Control) or 1 μM 4-OHT (n = 3–5). All statistical significances were calculated using two-tailed Student’s t-tests, except b where two-tailed Wilcoxon test was used. *p < 0.05; **p < 0.01; ***p < 0.001; ns non-significant. All error bars represent means ± s.d. Source data are provided as a Source Data file.
