Fig 1: Modulation of DNAJB4 protein level by m6A writer and erasers. a Western blot for monitoring the expression levels of DNAJB4, Flag-tagged FTO, and ALKBH5, and endogenous FTO in M14 cells transfected with control plasmid or plasmids for the ectopic expression of Flag-FTO or ALKBH5 at different time intervals following treatment with 100 nM ganetespib. b Quantification data based on western blot analysis in (a). c Western blot for monitoring the expression levels of DNAJB4 in M14 cells treated with control non-targeting siRNA (siCtrl), siMETTL3–1 or siMETTL3–2 at different time points following treatment with 100 nM ganetespib. d Quantification data based on western blot analysis in (c). β-actin was employed as the loading control in (a) and (c). Shown in (b) and (d) are the ratios of expression of DNAJB4 protein over β-actin, and further normalized to the ratios obtained for the control cells without ganetespib treatment. The quantification data in (b) and (d) represent the mean ± S. D. of results from three independent experiments. The p-values referred to comparisons between control cells and cells with ectopic overexpression of the indicated genes (b), or between controls and siRNA-mediated knockdown of the indicated genes (d). The p-values were calculated using unpaired, two-tailed Student’s t-test: #p > 0.05; *, 0.01 ≤ p < 0.05; **, 0.001 ≤ p < 0.01; ***, p < 0.001. Source data are provided as a Source Data file
Fig 2: HSP90 inhibitor-induced temporal changes in protein levels. Western blot images and the quantification data showing the alterations in expression levels of the HSP70, DNAJB4, METTL3, FTO, ALKBH5, and YTHDF3 in M14 cells at different time points following treatment with 100 nM ganetespib (a), AT13387 (b), or 17-DMAG (c). Shown are the ratios of expression of the indicated proteins over β-actin, and further normalized to the ratios obtained for the control cells without HSP90 inhibitor treatment. The data represent the mean ± S.D. of results from three independent experiments. The p-values referred to the comparison between 0 h and 24 h treatment and were calculated using unpaired, two-tailed Student’s t-test: #p > 0.05; *, 0.01 ≤ p < 0.05; **, 0.001 ≤ p < 0.01; ***, p < 0.001. Source data are provided as a Source Data file
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