Figure 4.

Gap junctional intercellular communication (GJIC) detected by the scrape loading and dye transfer (SLDT) method in Alveolar Type II (APTII) cells on day 4 of culture. Confluent monolayers were scraped with a sharp scalpel and loaded with Lucifer Yellow (LY, 457.25 kDa) (5% in PBS) for 5 min at room temperature. Observations were performed under epifluorescence. (a) Number of APTIIs from Cx43^+/+ mice stained in green was counted in each side of the rhodamine‐albumin line (red, 10 kDa) used as a negative control. (b) APTIIs from Cx43^+/+ mice. (c) APTIIs from Cx43^+/– mice. Note in the centre lane that rhodamine‐albumin remains on the scraped area. These experiments were repeated three times for each plate with similar results. For each experiment, 10⁷ APTII cells were plated initially. On day 4 of culture the cells reached more than 90% of confluence and the cells were scraped in region with complete confluence. *Represent significant results (P < 0.05).