No 66535 1 ig

Immunofluorescence was performed as previously described [17 , 20 (link)]. Briefly, after treatments, cells were fixed, permeabilized, and stained with primary antibodies against DHX9 (No.17721–1-AP, Proteintech), p65 (No.66535–1-Ig, Proteintech), p65 (phospho S536) (p-p65, sc-136548, Santa Cruz), F4/80 (No.28463–1-AP, Proteintech) at 4 °C overnight. After incubating with the second antibodies, cellular nuclei were stained with DAPI (0.5 μg/ml, Cell signaling) and then imaged by a Carl Zeiss LSM 800 confocal.

Western blotting was performed as previously described [17 , 20 (link)]. Briefly, cells were collected, lysed, and then quantified with a BCA kit (Beyotime, China). 20 μg of protein was loaded into a SDS gel, followed by electronic transfer onto PVDF membranes (Millipore, USA), and then incubated with primary antibodies against DHX9 (No.17721–1-AP, Proteintech), STAT1 (No.10144–2-AP, Proteintech), STAT1 phospho Ser727 (p-STAT1, No.39634, Active motif), p38 (No.14064–1-AP, Proteintech), Phospho-P38 MAPK (Thr180/Tyr182) (p-p38, No.28796–1-AP, Proteintech), JNK (No.66210–1-Ig, Proteintech), Phospho-JNK (Tyr185) (p-JNK, No.80024–1-RR, Proteintech), ERK1/2 (No.11257–1-AP, Proteintech), Phospho-ERK1/2 (Thr202/Tyr204) (p-ERK1/2, No.80031–1-RR, Proteintech), p65 (No.66535–1-Ig, Proteintech), p65 (phospho S536) (p-p65, sc-136548, Santa Cruz), GAPDH (ab8245, Abcam), flag (ab205606, Proteintech), or Lamin B1 (No.66095–1-Ig, Proteintech). After incubating with the secondary antibodies, the protein bands were detected using an ECL kit (Pierce, USA). The experiment was repeated three times independently.