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. 2022 Dec 12;24(2):e55472. doi: 10.15252/embr.202255472

Figure 3. LPS stimulation induces p38 MAPK‐dependent phosphorylation of TFEB‐S401 in macrophages.

Figure 3

  1. Immunoblot analysis of protein lysates from Raw 264.7 cells stably expressing TFEB‐WT‐FLAG incubated with 1 μg/ml LPS for the indicated times.
  2. Immunoblot analysis of protein lysates from Raw 264.7 cells stably expressing TFEB‐WT‐FLAG incubated with either 20 μM SB203580 or 250 nM Torin‐1 for 1 h prior to the addition of 1 μg/ml LPS for 30 min.
  3. Quantification of immunoblot data shown in (B). Data are presented as mean ± SD using one‐way ANOVA (unpaired) followed by Tukey's multiple comparisons test, (ns) not significant, *P < 0.05 and **P < 0.01 from three independent experiments.
  4. Immunoblot analysis of protein lysates from THP1 cells incubated with 1 μg/ml LPS for the indicated times.
  5. Immunoblot analysis of protein lysates from THP1 cells incubated with either vehicle (DMSO) or 20 μM SB203580 for 1 h prior to the addition of 1 μg/ml LPS for the indicated times.
  6. Immunoblot analysis of protein lysates from THP1 cells depleted of p38 MAPK (α), p38 MAPK (β) or p38 MAPK (α + β) and incubated with 1 μg/ml LPS for 1 h. Immunoblots are representative of at least three independent experiments.
  7. Quantification of immunoblot data shown in (F). Data are presented as mean ± SD using one‐way ANOVA (unpaired) followed by Tukey's multiple comparisons test, *P < 0.05 from three independent experiments.

Data information: n = 3 biological replicates (each dot represents a biological replicate).

Source data are available online for this figure.