Fig. 7: Inhibition of the MAPK pathway reduces FOSL1 level and sensitizes TEAD inhibitor resistant cell lines. | Nature Communications

Fig. 7: Inhibition of the MAPK pathway reduces FOSL1 level and sensitizes TEAD inhibitor resistant cell lines.

From: Cooperation between the Hippo and MAPK pathway activation drives acquired resistance to TEAD inhibition

Fig. 7

A Viability of H226-7883R cells measured after 4 days in presence of 1 μM GNE-7883, in the background of FOSL1 knockdown with three transfected individual guide RNAs, compared to the non-targeted control (NTC). Statistical significance was assessed using one-way ANOVA followed by posthoc Tukey’s test. Data are presented as mean values ± SD; n = 6 biological replicates. B H226-7883R cells were transfected with three single guide RNAs (sgRNA) targeting FOSL1 and control guide RNA targeting CD81, maintained in presence of GNE-7883 for 4 days, then fixed and stained with crystal violet solution to visualize cell colonies; n = 2 biological replicates. C Whole cell lysates of H226 and H226-7883R cells treated with DMSO, 1 μM GNE-7883 or Cobimetinib, analyzed by immunoblotting. (n = 3 biological replicates), Quantification in Fig. S11C. D Immunofluorescence of chromatin-bound FOSL1 in H226 and H226-7883R cells treated with DMSO, 1 µM GNE-7883 or 1 µM Cobimetinib for 48 h. Representative images for Hoechst and FOSL1 staining (Scale bar: 20 μm). The experiment was repeated three times with consistent results. E Boxplot of FOSL1 protein level distribution in the same cells as (D). n = 33491 for H226 treated with DMSO, 24102 for H226 treated with GNE-7883, 25875 for H226 treated with Cobimetinib, 37534 for H226-7883R treated with GNE-7883, 29316 for H226-7883R treated with Cobimetinib. The data are presented as medians ±25%, lower and upper hinges correspond to the 25th and 75th percentiles respectively. The lower and upper whiskers extend to the smallest and largest values within ±1.5 × the interquartile range from the hinges. Statistical analysis was performed using two-sided Wilcoxon rank-sum test on 500 randomly selected cells using datasample function in MATLAB. F Heatmaps showing the relative viability across a dose–response matrix of GNE-7883 and Cobimetinib combination (left) and GNE-7883 and Belvarafenib combination (right) in H226 and H226-7883R cells in 7-day viability assays. 1 indicates maximum viability and 0 indicates no viability. Measured IC50 for the indicated drug concentration is shown by the continuous line. Source data are provided as a Source Data file.

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