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Fig. 5 | BMC Biology

Fig. 5

From: TRIM25 promotes Capicua degradation independently of ERK in the absence of ATXN1L

Fig. 5

TRIM25 and ATXN1L mediated CIC stability in glioma. a Representative Western blot of CIC, ATXN1L, and phosphorylated ERK (pThr202/Tyr204) expression in GBM cell lines. b Representative Western blot of CIC, ATXN1L, and phosphorylated ERK (pThr202/Tyr204) expression in BTIC cell lines. c Tukey boxplots showing H-scores of CIC immunohistochemistry staining on glioma samples. d Immunohistochemistry images of CIC staining on glioma samples. Black bars denote 200 μm. e Representative Western blot of BTIC cell lines in standard EGF/FGF culture conditions and following 16 h of MEK/ERK inhibition. f Representative Western blot of BTIC cell lines following siRNA knockdown of ATXN1L or TRIM25. Fluorescent RNA was used as a negative control. g Relative mRNA expression of CIC and CIC target genes (DUSP6, SPRY4, ETV1/4/5) following treatment with ATXN1L or TRIM25 siRNA for 48 h in LN18 and U251 cell lines. Scrambled siRNA was used as a negative control. Gene expression was normalized to TBP. h Representative Western blot of LN229 and U343 cell lines treated with MEK/ERK inhibitors trametinib/LY3214996 and/or ATXN1L siRNA. DMSO and scrambled siRNA were used as a negative control. Below: barplot quantifications of CIC protein expression. i Representative Western blot of U251 and U343 cell lines treated with ATXN1L and/or TRIM25 siRNA. Scrambled siRNA were used as a negative control. Below: barplot quantifications of CIC protein expression. Western blot and RT-qPCR quantifications were collected from three independent experiments. Error bars represent one standard deviation. p values were calculated using the two-tailed independent Student’s t test. Statistically significant values are denoted (*p < 0.05, **p < 0.01). Individual data values can be found in Additional file 17: Table S10

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