Fig 9.

α-SMA expression in initial and 4-week-old cultures of HPTCs. (A) The expression levels of α-SMA (relative to GAPDH, average ± S.D.) were determined by qRT-PCR in initial cultures of HPTCs. These initial cultures contained cells freshly seeded from the vial obtained from the vendor, and the cells had not been passaged before analysis. The analysis was performed as soon as an epithelial sheet had been formed. For comparison, similar qRT-PCR analyses were also performed with confluent monolayer cultures of HEK293 and HeLa cells, and the results are shown. (B) α-SMA expression (relative to GAPDH, average ± S.D.) was determined by qRT-PCR in initial cultures of HPTCs (day 0) and 28 days later in cultures that were seeded in parallel. The cultures were not passaged during this time period, but the medium was regularly exchanged. (C) The image shows an initial culture of HPTCs (day 0) after co-immunostaining (ZO-1: green, α-SMA: red, DAPI: blue). α-SMA was not detectable. (D) The same co-immunostaining procedure was performed after 28 days with cultures seeded in parallel. Many α-SMA-expressing cells are present.