Fig. 5. UHMK1 associates with mRNA encoding metabolic proteins and promotes selective mRNA transport in BRAF^V600 melanoma cells adapting to BRAFi.

A375 cells were transfected with the indicated siRNA and treated with DMSO or 1 μM Vem for 48 h. a RNA fluorescence in situ hybridization (FISH) using a poly(A)⁺ RNA specific probe in A375 cells treated as indicated (representative of n = 3 biologically independent experiments). b The nuclear to cytoplasm ratio of poly(A)⁺ RNA was calculated using high content image analysis. Data are expressed as mean fold change ± SEM (n = 3 biologically independent experiments). Statistical significance was determined using a one-way ANOVA adjusted for multiple comparisons. c RNA immunoprecipitation (RNA-IP) assays were performed in UHMK1-V5 expressing A375 cells following treatment with DMSO or 1 μM Vem for 48 h. The indicated mRNA transcripts were then analysed using RT-qPCR. Data are expressed as mean ± SEM (n = 3 biologically independent experiments). Statistical significance was determined using a one-way ANOVA adjusted for multiple comparisons. d Cell lysates were fractionated into nuclear and cytoplasmic pools of RNA and analysed for the indicated genes using RT-qPCR. The nuclear/cytoplasm (Nuc/Cyto) ratio was calculated from analysis of individual nuclear and cytoplasmic compartments. Data are expressed as mean ± SEM (n = 3 biologically independent experiments). See also Supplementary Fig. 5C. Statistical significance was determined using a one-way ANOVA adjusted for multiple comparisons. e Whole-cell lysates were used to assess total mRNA levels. Data are expressed as mean ± SEM (n = 2 biologically independent experiments). See also Supplementary Fig. 5. Source data are provided as a Source Data file.