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. 2022 Aug 2;11:e80725. doi: 10.7554/eLife.80725

Figure 5. Glutamine deprivation modulates mTOR activation.

(A) Primary murine chondrocytes were cultured in media containing 4 mM glutamine or 0 mM glutamine for 24 hr. Cells were then treated with IL-1β (10 ng/mL). After 24 hr, lysates were collected, and immunoblotting was performed for pAKT (60kDa) and total Akt (58kDa). (B) Under similar conditions, immunoblotting was performed for pS6 and total S6 (32kDa). (C–D) Primary murine chondrocytes were cultured in media containing 4 mM glutamine or 0 mM glutamine for 24 hr. Cells were then treated with IL-1β (10 ng/mL) in the presence or absence of rapamycin 50 nM for 24 hr. Gene expression of Il6 and Mmp13 was measured by quantitative PCR. Results from one representative experiment. n=4. One-way ANOVA was performed followed by Tukey’s multiple comparisons test. C:****p<0.0001.

Figure 5—source data 1. Depicting original raw data related to Figure 5.
Figure 5—source data 2. Original raw data related to Figure 5A, B.

Figure 5.

Figure 5—figure supplement 1. Glutamine deprivation modulates mTOR activation.

Figure 5—figure supplement 1.

(A) Primary murine chondrocytes were cultured in media containing 4 mM glutamine or 0 mM glutamine for 24 hr. Cells were then treated with IL-1β (10 ng/mL) for 24 hr in the presence or absence of rapamycin (50 nM). Immunoblotting was performed for pS6 and total S6 (32kDa). (B–E) Under similar conditions, gene expression was measured by quantitative PCR. One-way ANOVA was performed followed by Tukey’s multiple comparisons test. B:****p<0.0001, C: **p=0.0017, and D:**p=0.0088. n=4.
Figure 5—figure supplement 1—source data 1. Depicting original raw data related to Figure 5—figure supplement 1.
Figure 5—figure supplement 1—source data 2. Original raw data related to Figure 5—figure supplement 1A.