Journal: PLoS ONE

Article Title: Alpha-Synuclein Proteins Promote Pro-Inflammatory Cascades in Microglia: Stronger Effects of the A53T Mutant

doi: 10.1371/journal.pone.0162717

Figure Lengend Snippet: After 30 min, 1 h, 3 h or 6 h of A53T protein exposure (5 μM), microglial cell lysates were subjected to western-blot analysis (Fig 5A) to determine the expression of phospho-Stat1 (pStat1), phospho-p38 (pp38) and phospho-ERK (pERK). α-Tubulin (αTub) was used as a loading control. These chemiluminescent detection assays were realized with 20 μg of total proteins. Ratios between phosphorylated versus non-phosphorylated proteins, called pStat1/Stat1, pp38/p38 and pERK/ERK, were described in Fig 5B. Results are given as mean ± SEM of at least three independent experiments. * p < 0.05, significantly different from control condition.

Article Snippet: Blots were incubated with a mouse anti-Stat1 (1:500, BD Biosciences, Belgium), rabbit anti-phospho-Stat1 (1:1000, Bioke, The Netherlands), rabbit anti-p38 (1:1000, Calbiochem, Belgium), rabbit anti-phospho-p38 (1:1000, Bioke), rabbit anti-ERK (1:20000, Sigma), rabbit anti-phospho-ERK (1:2000, Bioke), mouse anti-α-tubulin (1:5000, AbCam, UK), rabbit anti-p65 (1:200, Santa Cruz Biotechnologies, Germany), rabbit anti-c-Fos (1:1000, Bioke), rabbit anti-Nrf2 (1:1000, Bioke) and mouse anti-HDAC1 (1:2000, AbCam) antibody.

Techniques: Western Blot, Expressing