Fig. 2: Effect of TMEM45A inactivation on cell morphology and proliferation in SQD9 and RCC4 cells.

SQD9 and RCC4 cells were transduced with lentiviral particles expressing shRNA control (shCTL) or shRNA targeting the mRNA of TMEM45A (shRNA22) or were not transduced (UT). a SQD9 cell morphology was studied by optical microscopy (magnification ×200) and b transmission electron microscopy. c RCC4 cell morphology was studied by optical microscopy (magnification ×200). d, e 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 and gene expression level was assessed by RNA sequencing. d Ten first genes differentially expressed in the knockdown cells in the absence or in the presence of cisplatin. e 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 G2/M transition pathway. 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. f After the tranduction, SQD9 cells were incubated in normoxic conditions for 24 and 48 h then, the proliferation was assessed by MTT assay. The signal intensity of the control cells at the seeding (time 0) was used to normalize the number of viable cells at different time points. Results are expressed as mean ± SD (n = 3). **p < 0.01. g After the transduction, RCC4 cells were incubated in normoxic conditions for 24, 48, and 72 h then, the proliferation was assessed by MTT assay. The signal intensity of the control cells at the seeding (time 0) was used to normalize the number of viable cells at different time points. Results are expressed as mean ± SD (n = 3). N normoxia, NC normoxia + cisplatin. **p < 0.01.