Fig. 4

Microglial DICER is phosphorylated at serine 1456 by JNK. a Representative immunoblots of immunoprecipitates from the total lysates of HEK293 cells transfected with control (Ctrl), JNK, or/and HA-tagged DICER (HA-DICER) constructs and probed with anti-phospho Ser antibody. Lower—immunoblots of the precipitates with anti-HA antibody. b, c Representative immunoblots of immunoprecipitates of DICER from total lysates of microglia (b) and BV2 cells (c) exposed to MPP⁺ for 30 min and probed with the anti-phospho Ser antibody. d Upper—representative immunoblots of immunoprecipitates from total lysates of BV2 cells preincubated with SP600125 for 2 h then with MPP⁺ for 30 min and probed with the anti-phospho Ser antibody. Lower—statistics (n = 3). Data are shown as mean + SEM. *p < 0.05. e Mass spectrometry (MS) analysis of the DICER immunoprecipitated from the microglia incubated in the absence (upper, Ctrl) or presence (lower, MPP⁺) of MPP⁺ for 30 min. The phosphorylated serine residue was shown in lowercase bold letter. f Multiple sequence alignment of DICER containing the conserved serine residue in Mus musculus (Mm) (1444–1469), Rattus norvegicus (Rn) (1456–1481), and Homo sapiens (Hs) (1458–1483). Residues are colored based on percentage identity