Figure S4.
CM augments Cer synthesis–related gene transcription in receiving cells, and CRT reduces the content of Cer in cells. (A and B) Quantification of protein levels in Fig. 7 A (A) and Fig. 7 B (B). Lanes were numbered from left to right, with the ratio in lane 2 (Tg treatment) set as 100. The statistical analyses were calculated by one-way ANOVA. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (C) Cer concentration in 3T3-L1 and AML12 treated without or with 3T3-L1–derived CM. Left panel, Cer species that showed no significant difference of the concentration between 3T3-L1 and AML12 (unpaired t test, two-tailed, P ≥ 0.05). Right upper panel, Cer species that showed a significant difference in the concentration between 3T3-L1 and AML12 (unpaired t test, two-tailed, P < 0.05). Right lower panel, total Cer. The statistical analysis between AML12 and AML12 treated with CM was calculated using an unpaired t test, two-tailed. Data were shown as the mean ± SEM, n = 4. *P < 0.05. (D) RT-qPCR of Cer synthesis–related genes in AML12 treated with Ctrl/CM for 30 min. The statistical analyses were calculated by an unpaired t test, two-tailed. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001. (E) Quantification of protein levels in Fig. 7 J. Lanes were numbered from left to right, with the ratio in lane 2 (Tg treatment) set as 100. The statistical analyses were calculated by an unpaired t test, two-tailed. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01. (F) Representative confocal microscopic images of AML12 cells stained with anti-Cer antibody and Hoechst (blue). Cells were treated with Ctrl/CM ± 5 μM CRT0066101 (CRT) for 2 h. Scale bar, 20 μm. (G) Quantification of Cer fluorescence intensities in F measured by ImageJ, n ≥ 8 fields. The statistical analyses were performed using one-way ANOVA. Data were shown as the mean ± SEM. *P < 0.05, ***P < 0.001. (H) Quantification of protein levels in Fig. 7 K. Lanes were numbered from left to right, with the ratio in lane 2 (Tg treatment) set as 100. The statistical analyses were calculated by one-way ANOVA. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01. Source data are available for this figure: SourceData FS4. Refer to the image caption for details.

CM augments Cer synthesis–related gene transcription in receiving cells, and CRT reduces the content of Cer in cells. (A and B) Quantification of protein levels in Fig. 7 A (A) and Fig. 7 B (B). Lanes were numbered from left to right, with the ratio in lane 2 (Tg treatment) set as 100. The statistical analyses were calculated by one-way ANOVA. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (C) Cer concentration in 3T3-L1 and AML12 treated without or with 3T3-L1–derived CM. Left panel, Cer species that showed no significant difference of the concentration between 3T3-L1 and AML12 (unpaired t test, two-tailed, P ≥ 0.05). Right upper panel, Cer species that showed a significant difference in the concentration between 3T3-L1 and AML12 (unpaired t test, two-tailed, P < 0.05). Right lower panel, total Cer. The statistical analysis between AML12 and AML12 treated with CM was calculated using an unpaired t test, two-tailed. Data were shown as the mean ± SEM, n = 4. *P < 0.05. (D) RT-qPCR of Cer synthesis–related genes in AML12 treated with Ctrl/CM for 30 min. The statistical analyses were calculated by an unpaired t test, two-tailed. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001. (E) Quantification of protein levels in Fig. 7 J. Lanes were numbered from left to right, with the ratio in lane 2 (Tg treatment) set as 100. The statistical analyses were calculated by an unpaired t test, two-tailed. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01. (F) Representative confocal microscopic images of AML12 cells stained with anti-Cer antibody and Hoechst (blue). Cells were treated with Ctrl/CM ± 5 μM CRT0066101 (CRT) for 2 h. Scale bar, 20 μm. (G) Quantification of Cer fluorescence intensities in F measured by ImageJ, n ≥ 8 fields. The statistical analyses were performed using one-way ANOVA. Data were shown as the mean ± SEM. *P < 0.05, ***P < 0.001. (H) Quantification of protein levels in Fig. 7 K. Lanes were numbered from left to right, with the ratio in lane 2 (Tg treatment) set as 100. The statistical analyses were calculated by one-way ANOVA. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01. Source data are available for this figure: SourceData FS4.

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