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Fig. 3 | BMC Biology

Fig. 3

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

Fig. 3

ERK activity relieves CIC’s repressive function. a Representative Western blot of ATXN1LWT (HEK) and ATXN1LKO (A30) cell lines treated with FGF/EGF over 0–24 h following serum starvation. FBS control was cultured in FBS for the duration of the time course. Below: barplot quantifications of CIC expression. Quantifications were normalized to vinculin. b Representative Western blot of ATXN1LWT (HEK) and ATXN1LKO (A30) cell lines treated with FGF/EGF and/or MEK/ERK inhibitors trametinib/LY3214996. c Immunofluorescence images of NHA cells cultured in FBS, serum starvation, and EGF/FGF. White bars denote 10 μm. d Relative mRNA expression of ATXN1L, CIC, and CIC target genes DUSP6, SPRY4, and ETV1/4/5 in ATXN1LWT (NHA) and ATXN1LKO (B82) cell lines treated with FGF/EGF for 8 h following serum starvation. Gene expression was normalized to TBP, and the serum-starved parental ATXN1LWT (NHA) cell line was used as a relative control. e ChIP-PCR showing relative enrichment of the CIC-DNA interaction following serum starvation or treatment with EGF/FGF (1 h) in ATXN1LWT (HEK) and ATXN1LKO (A30) cell lines. Relative enrichment was normalized to a control region (NCR). Western blot, RT-qPCR, and ChIP-PCR quantifications were collected from 3 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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