Fig. 6: Effect of TMEM45A inactivation on UPR pathway activation.

a, b SQD9 cells were transduced with lentiviral particles expressing shRNA control (shCTL) or shRNA targeting the mRNA of TMEM45A (shRNA 22). The cells were incubated with or without 100 mM of cisplatin in normoxic (N) conditions for 24 h. (A) Genesexpression level was assessed by RNA sequencing. The transcriptome analysis was performed using Babelomics and GSEA after RNA sequencing. Heatmap of differentially expressed genes produced by using the heatmapper²⁶ (http://www.heatmapper.ca) showed a potential deregulation of the Unfolded Protein Response activation. Genes have been reordered by the method of average clustering. A dendrogram is shown for the clustering of genes. Genes with relatively high expression are marked in red, genes with relatively low expression are marked in blue. The corresponding values are presented in Supplementary Fig. 4. b After mRNA extraction the expression level of several target genes of UPR pathway was assessed by RT-qPCR. c SQD9 cells were incubated with or without 20 mM of cisplatin in normoxic conditions for 48 h. Splicing of XBP1 was assessed by PCR and migration on agarose gel. The signal intensity was measured by Image J. Results are expressed as mean ± SD (n = 3). d, e RCC4 cells were transduced with lentiviral particles expressing shRNA control (shCTL) or shRNA targeting the mRNA of TMEM45A (shRNA 22). RCC4 cells were incubated with or without 20 mM of cisplatin in normoxic conditions for 48 h. d After mRNA extraction the expression level of several target genes of UPR pathway was assessed by RT-qPCR. e Splicing of XBP1 was assessed by PCR and migration on agarose gel. The signal intensity was measured by Image J. Results are expressed as mean ± SD (n = 3). N normoxia, NC normoxia + cisplatin. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.