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Fig. 2 | Journal of Translational Medicine

Fig. 2

From: LINC00665 knockdown confers sensitivity in irradiated non-small cell lung cancer cells through the miR-582-5p/UCHL3/AhR axis

Fig. 2

UCHL3 maintained the stability of AhR protein through deubiquitination, thereby promoting PD-L1 expression. A, Positive expression of AhR protein in tumor tissues and adjacent normal tissues from NSCLC patients (n = 45) detected by Immunohistochemistry. B, Silencing efficiency of sh-UCHL3 in A549 cells and overexpression efficiency of LV-UCHL3 in PC9 cells detected by qRT-PCR. C, Protein levels of UCHL3 and AhR in A549 cells treated with sh-UCHL3 or in PC9 treated with LV-UCHL3 detected by Western blot analysis. D, Western blot analysis of AhR protein degradation in A549 and PC9 cells following CHX (10 μg/mL) treatment (the left penal) and corresponding statistical analysis of AhR protein stability (the right penal). E, Co-IP detection of exogenous UCHL3 and AhR proteins in HEK293T cells. F-G, AhR protein degradation rate in A549 cells in response to sh-UCHL3 (F) or in PC9 cells in response to LV-UCHL3 (G) detected by CHX pulse-chase analysis. H, Detection of AhR protein ubiquitination in A549 cells overexpressing UCHL3 or in PC9 cells silencing UCHL3. I, Expression of PD-L1 in tumor tissues and adjacent normal tissues from NSCLC patients (n = 45) detected by qRT-PCR. J, Expression of PD-L1 in A549 cells treated with sh-UCHL3 or in PC9 cells treated with LV-UCHL3 detected by qRT-PCR. K, Silencing efficiency of sh-AhR in A549 cells and overexpression efficiency of LV-AhR in PC9 cells detected by qRT-PCR (the left two panels) and Western blot analysis (the right two panels). L, Expression of PD-L1 in A549 cells treated with sh-AhR or in PC9 cells treated with LV-AhR detected by qRT-PCR. M, Protein levels of UCHL3, AhR and PD-L1 in A549 cells treated with sh-UCHL3 or combined with LV-AhR or in PC9 cells treated with LV-UCHL3 or combined with sh-AhR detected by Western blot analysis. * p < 0.05 versus the adjacent normal tissues or A549 cells treated with sh-NC; # p < 0.05 versus the PC9 cells treated with Vector; & p < 0.05 versus the A549 cells treated with sh-UCHL3 + Vector; @ p < 0.05 versus the PC9 cells treated with LV-UCHL3 + sh-NC. Each cellular experiment was repeated 3 times

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