Polymerase chain reaction (PCR)

Total RNA was extracted using TRIzol reagent (Life Technologies). First-strand cDNA was synthesized using Superscript III Reverse Transcriptase (Life Technologies). Real time quantitative PCR (qPCR) was performed in triplicates using SYBR green PCR master mix and a 7900HT Fast Real Time PCR system (Applied Biosystems). Products were analyzed with ABI 7900HT Sequence Detection System (Applied Biosystems) or Rotor gene (Corbett). 2^-ΔCt values are used to calculate the relative expression levels of 9 VDCC subtypes, with TATA box binding protein (TBP) as reference gene (S2 Table). For quantification of Ca_v1.3 knock-down, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and actin were used as reference genes (S2 Table). For quantification of T. gondii in tissues, the B1 gene was used (S2 Table).