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. 2022 May 3;13(5):427. doi: 10.1038/s41419-022-04857-y

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© The Author(s) 2022, corrected publication 2022

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Fig. 6 — A Real-time qRT-PCR assay indicating the expression of RORA in A549 cells stably transfected with empty vector (mock) or POU6F1. B Co-IP and western blotting assays indicating the endogenous interaction between POU6F1 and RORA in A549 cells. The immunoglobulin G (IgG)-bound protein was taken as the negative control. C Mass spectrometry detection showing the spectrogram of RORA in A549 and HEK293T cells pulled down by POU6F1 antibody. D Immunofluorescence assay confirming the colocalization of POU6F1 and RORA in A549 cells transfected with empty vector (pCMV-HA) or pCMV-HA-POU6F1. Scale bars:10 µm. E Confocal images of BiFC showing the direct visualization of POU6F1 and RORA in A549 cells co-transfected with pBiFC-VC155-POU6F1 and pBiFC-VN173-RORA. Scale bars:10 µm. F Western blotting assay showing the expression of RORA in A549 and NCI-H1299 cells transfected with mock, POU6F1, CRISPRi-Scb, CRISPRi-POU6F1 #1, or CRISPRi-POU6F1 #1. G Co-IP and western blotting assays indicating the interaction between POU6F1 and RORA in A549 cells transfected with mock or POU6F1. H Western blotting assay showing the expression of POU6F1 and RORA in A549 cells transfected with mock, RORA, scramble shRNA (sh-Scb), sh-RORA #1, or sh-RORA #2. I Western blotting assay showing the expression of RORA in A549 cells transfected with CRISPRi-Scb or CRISPRi-POU6F1 #1, and those treated with MG132 (5 μmol/l) for 6 h. J Dual-luciferase assay indicating relative promoter activity of RORA in A549 and NCI-H1299 cells transfected with mock or POU6F1. Student t-test compared the difference in A and J. *P < 0.05, **P < 0.01.