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. 2023 Apr 6;18(4):e0283894. doi: 10.1371/journal.pone.0283894

Fig 6. Immunoblot analysis and IF of 3xFLAG RNF43 knockin clones.

Fig 6

A. Immunoblot analysis of Caco-2 and OE19 clones with a 3xFLAG knockin at the C-terminus of RNF43. A sensitive ECL Ultra Western HRP Substrate system was required to reveal the bands. A 30x diluted sample of RNF43-FLAG transfected HEK293T cells is added as control. B. Using a Tyramide SuperBoost system a small number of 3xFLAG positive OE19 cells could be identified. No positive cells were observed in the parental OE19 cell line. The staining pattern resembles that of overexpressed RNF43 showing a cytoplasmic and perinuclear pattern, the latter reminiscent of an ER location. However, given the low number of positive cells in only one cell line, we cannot draw a reliable generalized conclusion about RNF43’s intracellular location. Scale bar, 25um.