Pet28a expression vector

Coding sequences were amplified from Botrytis cinerea cDNA or plasmid DNA. Recipient vectors and fragments were cut using indicated restriction sites (Table S3). Vectors were dephosphorylated prior ligation and fragment purified via gel-extraction if necessary. For transient expression, pGreen II derivatives (pFF04 and pFF18) were used as recipient vectors (Künzl et al., 2016) . Both vectors harbour a cauliflower mosaic virus (CMV) 35S promotor, an N-terminal signal peptide (SP) and a kanamycin resistance cassette. A hemagglutinin (HA)tag for immunological detection was added at the C-terminus of the coding sequence via the reverse primers. The Hip1-GFP fusion was constructed by GreenGate cloning (Lampropoulos et al., 2013) , using the following modules: 35S promotor (pGGA004), ER signal sequence (pGGB006), linker-GFP (pGGD001) and RBCS terminator (pGGE001) The Hip1 coding sequence devoid of BsaI restriction sites for GreenGate cloning was synthesized by Biocat (Germany). For recombinant protein expression, the Hip1 sequence was amplified without signal peptide and cloned into the pET28a (+) expression vector (Addgene, USA), eventually having a His-tag at the N-terminus. All primers used are shown in Table S3. Gene constructs were verified by sequencing (Seq-It, Germany). For colocalization studies, the PM marker spRFP-TMD23 (pFK44) was used (Scheuring et al., 2012) .