Figure 5
pH e effect on prolactin signaling is rapidly reversible. (A) SKBR3 cells were preincubated with medium of either pHe 7.4 or pHe 6.8 for 30 min at 37°C. Then the pHe was altered or not as indicated and cells were treated with prolactin for 15 min. Two groups were incubated further for another 30 minutes as indicated before exposure to prolactin. Jak2, Stat5 and Erk activation was analyzed (n = 3). (B) T47D spheroids grown in three-dimensional culture conditions were treated with prolactin or vehicle at indicated pH conditions. The spheroids were stained for pYStat5. The experiment was carried out twice.