No statistical methods were used to determine sample size. Experiments were not randomized, and investigators were allocated without blinding during experimentation and analyses. All experiments were repeated at least twice. For ELISA, quantitative reverse transcription PCR, LDH release, and PI uptake assays, two to three technical replicates were used to estimate experimental mean. Means from three or more biologically independent experiments, as indicated by the values of n in legends, were analyzed by statistical methods. For immunofluorescence assays, typically ~50 to 100 host cells or 100 to 200 L. monocytogenes were counted and mean percentage of cells showing events were obtained, and mean percentage from three biologically independent repeats were compared. Data were normally distributed (based on D’Agostino-Pearson or Shapiro-Wilk normality tests) after logarithm transformation and were then analyzed by statistical methods. Repeated-measures ANOVA was used to analyze means from immunofluorescence assays and immunoblot quantifications, and paired two-tailed Student’s t test was used elsewhere. When more than three comparisons were made, P values were adjusted for multiple comparisons by the Benjamini-Krieger-Yekutieli method (Q = 0.05, false discovery rate) implemented in GraphPad Prism 7. Discoveries at q < 0.05 are reported. Unless otherwise indicated, means ± SEM are plotted. L. monocytogenes ΔactA CFU assays with Δp62 cells were performed six times alongside +WT and +p62TRM cells, and +D329G and +D329H cell lines were also included in five experiments; these data are shown as matched means in Figs. 3F and 5D. In IL-6 ELISAs shown in Figs. 4C, 5E, and 7B, every experiment included Δp62 and typically also +WT cell line, alongside the indicated p62 variants; means ± SEM from all experiments are plotted, and data for Δp62 cells are shown in each figure for comparison. Data plots and statistics used Prism (version 7.04, GraphPad Software Inc.).

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