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. 2019 Oct 25;13:468. doi: 10.3389/fncel.2019.00468

Figure 3.

Figure 3

Inhibition of HDAC4 transcriptionally downregulated MKK7 expression and lowered 5K-evoked JNK/c-Jun activity. (A) Human glioma cell line U251 was transfected with siNC or the small interference RNAs (siRNAs) against indicated HDAC members (Supplementary Table S1), and MKK7 mRNA levels were detected by Q-PCR. Mean ± SD, *P < 0.05 vs. siNC. (B) Rat C6 glioma cells were transfected with siNC, siHDAC4-a, or siHDAC4-b, and mkk7 mRNA or MKK7 protein levels were determined by Q-PCR or WB, respectively. Mean ± SD, *P < 0.05 vs. siNC. (C) DIV5 CGNs transfected with siNC, siHDAC4-a, or siHDAC4-b together with pGFP plasmids were subjected to IF for HDAC4 and MKK7 co-staining. Photos were obtained using a confocal microscope (scale bar = 20 μm). GFP was used to mark the transfected cells, and the effect of HDAC4 knockdown on MKK7 expression was determined by scoring the percentage of the GFP-positive neuron population with HDAC4 or MKK7 stained. Mean ± SD, *P < 0.05 vs. siNC, #P < 0.05 vs. siNC. (D–F) CGNs in 25K or 5K media that contained LMK235 at the indicated doses for 12 h were collected to detect the levels of MKK7, p-JNK, p-c-Jun, c-Jun by WB, or mkk7 mRNA by Q-PCR. Mean ± SD, *P < 0.05 vs. siNC, #P < 0.05 vs. 0.1 μM LMK235.