Fig. 7: LARP6 NTR is dispensable for RNA localisation, but critical for regulation of cancer cell viability and invasion.
From: An intrinsically disordered region mediates RNA-binding selectivity and cellular activities of LARP6
a Experimental workflow for profiling RNA localisation to protrusions and cell bodies of U-87 MG cells, using 3 µm transwell filters. Cells were seeded on top of filters and allowed to form protrusions through the pores for 90 minutes. RNA was isolated from each side of the filter and subjected to 3’ end RNA-sequencing to quantify relative mRNA enrichments in protrusions vs. cell-bodies (Created with modifications, using BioRender. Mardakheh, F. (2026) https://BioRender.com/16v6db4). b Profile plot of log2 protrusions/body ratio values for all ribosomal protein encoding mRNAs, in the indicated U-87 MG cells (n = 3 biological replicates). Positive values indicate enrichment in protrusions (blue arrow), while negative values indicate enrichment in cell bodies (red arrow). The boxplots mark the lower and upper quartiles with the median marked by the horizontal line. The whiskers extend to the minima and maxima, excluding outliers. An unpaired two-sided two-sample t-test was used for the calculation of significance. P values are indicated on the bar graphs. c Normalised invasion index for the indicated U-87 MG cell populations upon endogenous LARP6 knockdown and rescue with doxycycline-inducible myc-LARP6 FL or ΔNTR. The values were normalised to the average non-targeting parental control condition. Error bars = mean ± SD (n = 30 (Parentals), 31 (FL), 28 (ΔNTR) biological replicates, pooled from 3 separate experiments). An unpaired two-sided two-sample t-test was used for the calculation of significance. The p-values are indicated on the bar graphs. d Normalised cell viability for the indicated U-87 MG cell populations, upon endogenous LARP6 knockdown and rescue with doxycycline-inducible FL or ΔNTR myc-LARP6. Cell viability was assessed five days after siRNA transfection, using CTG assay. The values were normalised to the average non-targeting parental control condition. Error bars = mean ± SD (n = 14 (Parental), 16 (FL), 12 (ΔNTR) biological replicates, pooled from 3 separate experiments). A two-sided Kolmogorov–Smirnov test was applied for the significance calculation. The p-values are indicated on the bar graphs.
