Figure S7.
Synergy between NBR1 and TAX1BP1. (A) HeLa PentaKO cells were co-transfected with tf-NBR1, HALO-TAX1BP1, and the indicated BFP-tagged receptors. Triple-positive cells were analyzed by flow cytometry with red:green ratio as a readout of NBR1 flux. Graphs represent mean ± SD from three independent experiments. n > 10,000 cells per sample. P values were determined using a one-way ANOVA (P = 0.0006). Multiple comparisons were made using Dunnett T3 correction. **, P < 0.01, ns, not significant. (B) HeLa pentaKO cells were transfected with tf-TAX1BP1 (WT or R441E). After 24 h, cells were analyzed for red:green ratio by flow cytometry (n > 10,000 cells per sample). Median values for each sample are identified by a black line within each violin. BafA1, Bafilomycin A1. (C)Atg7KO/TAX1BP1KO K562 cells expressing tf-NBR1 were electroporated with the indicated BFP-tagged variants of TAX1BP1. BFP-expressing cells were analyzed by red:green ratio as a readout for flux. n > 10,000 cells per sample. (D) HeLa pentaKO cells were co-transfected with tf-NBR1 and the indicated BFP-tagged TAX1BP1 variants. tf-positive cells were analyzed by flow cytometry with red:green ratio as a readout of NBR1 flux. Graphs represent mean ± SD from three independent experiments. n > 10,000 cells per sample. P values were determined using a one-way ANOVA (P < 0.0001). Multiple comparisons were made using Tukey’s correction. ****, P < 0.0001, ns, not significant. Refer to the image caption for details.

Synergy between NBR1 and TAX1BP1. (A) HeLa PentaKO cells were co-transfected with tf-NBR1, HALO-TAX1BP1, and the indicated BFP-tagged receptors. Triple-positive cells were analyzed by flow cytometry with red:green ratio as a readout of NBR1 flux. Graphs represent mean ± SD from three independent experiments. n > 10,000 cells per sample. P values were determined using a one-way ANOVA (P = 0.0006). Multiple comparisons were made using Dunnett T3 correction. **, P < 0.01, ns, not significant. (B) HeLa pentaKO cells were transfected with tf-TAX1BP1 (WT or R441E). After 24 h, cells were analyzed for red:green ratio by flow cytometry (n > 10,000 cells per sample). Median values for each sample are identified by a black line within each violin. BafA1, Bafilomycin A1. (C)Atg7KO/TAX1BP1KO K562 cells expressing tf-NBR1 were electroporated with the indicated BFP-tagged variants of TAX1BP1. BFP-expressing cells were analyzed by red:green ratio as a readout for flux. n > 10,000 cells per sample. (D) HeLa pentaKO cells were co-transfected with tf-NBR1 and the indicated BFP-tagged TAX1BP1 variants. tf-positive cells were analyzed by flow cytometry with red:green ratio as a readout of NBR1 flux. Graphs represent mean ± SD from three independent experiments. n > 10,000 cells per sample. P values were determined using a one-way ANOVA (P < 0.0001). Multiple comparisons were made using Tukey’s correction. ****, P < 0.0001, ns, not significant.

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