Phospho-ALK (Tyr1507) Antibody - #AF3489

Fig. 3. Phosphorylated-ALK is involved in the S. pneumoniae infection-induced NLRP3 inflammasome activation. A-B: PECs were infected with S. pneumoniae at an MOI of 5 for 0, 5, 15, 30 or 60 min. A: The cell lysates were collected, and total and phosphorylation levels of ALK were detected by western blot. B: P-ALK/t-ALK was analyzed by relative densitometric quantification. C-J: PECs were pretreated with or without ALK inhibitor (Bri, 20nmol/L) for 2 h and then infected with S. pneumoniae (MOI of 5) for 6 h. C-F: The protein expression levels of pro-caspase-1, p20, pro-IL-1β and p17 in supernatants or cell lysates were detected by western blot and analyzed by relative densitometric quantification. G: The mRNA expression levels of Caspase-1 and Il-1β were quantified by RT-PCR. H: The protein expression level of NLRP3 were detected by western blot and analyzed by relative densitometric quantification. I: The mRNA expression level of Nlrp3 was detected by RT-PCR. J: The ASC specks formation were imaged via fluorescence microscopy. (Scale bar: 100 µm.) K: PECs were pretreated with or without Bri for 2 h and then infected with S. pneumoniae (MOI of 1) for 24 h.The supernatants were collected and the level of IL-1β in supernatants were detected by ELISA. L-N: PECs were pretreated with or without Bri for 2 h and then infected with S. pneumoniae (MOI of 5) for 6 h. L: The protein expression level of NEK7 were detected by western blot and analyzed by relative densitometric quantification. M: The mRNA expression level of Nek7 was detected by RT-PCR. N: The NEK7-NLRP3 complex formation were detected by Co-IP. Similar results were obtained in three independent experiments in data. Data shown are means ± SEMs; *P