Fig. 2: NLRP6 promotes p85α autophagic degradation via the cargo receptor OPTN.

a p85α protein (top) and PIK3R1 mRNA (bottom) levels were analysed in LN229 cells transfected with increasing amounts of Flag-NLRP6. p85α protein levels were quantified. b p85α protein (top) and PIK3R1 mRNA (bottom) levels were analysed in Control (Ctrl) and NLRP6 knockout (KO) LN229 cells. p85α protein levels were quantified. c LN229 cells transfected with Flag-NLRP6 were treated with DMSO (Mock), MG132, 3-methyladenine (3-MA), bafilomycin A1 (BafA1), chloroquine (CQ), or NH₄Cl, and the effects on p85α protein levels were examined. d p85α protein levels were analysed in Ctrl and ATG5 KO LN229 cells and were quantified. e Coimmunoprecipitation (Co-IP) analysis to investigate the interaction between p85α and LC3 in the presence of NLRP6 in HEK293T cells. WCL, whole cell lysates. f Co-IP analysis of the interaction between endogenous p85α and LC3 in the absence of NLRP6 in LN229 cells. g Representative confocal microscopy of LN229 cells transfected with NLRP6. Scale bar = 5 μm (top). Statistical analysis of the number for p85α and LC3 colocalization (bottom). h Co-IP analysis to map the interaction between p85α and cargo receptors in HEK293T cells. i Co-IP analysis of the interaction between p85α and OPTN in the absence of NLRP6 in LN229 cells. j p85α protein levels were analysed in Ctrl and OPTN KO LN229 cells transfected with Myc-NLRP6 (left) and were quantified (right). In a, b, d and j, all error bars, mean values ± SD, p-values were determined by unpaired two-tailed Student’s t test of n = 3 independent biological experiments. For g, error bar, mean value ± SD, p-value were determined by unpaired two-tailed Student’s t test of n = 19 cells. For c, e, f, h, and i, data shown are representative of three independent experiments with similar results. Source data are provided as a Source Data file.