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. 2019 Jul 22;11(8):e9830. doi: 10.15252/emmm.201809830

Figure 4. TGFβ/ActivinB signaling promotes cell proliferation in Group 3 MB cell lines.

Figure 4

  • A–H
    1603MED (A‐D) or D283 (E‐H) cells were treated with DMSO (vehicle, black), with LY364947 (red), or with SB431542 (orange). (A and E) Immunoblot of phosphorylated Smad2 (P‐Smad2), total Smad2, and β‐actin upon inhibition of TGFβ/Activin signaling using LY364947 and SB431542 inhibitors for 24 h. Bar graphs on the right panel represent the quantification of the relative level of P‐Smad2 (P‐S2) to β‐actin. (B and F) P‐Smad2 to total Smad normalization is provided on Appendix Fig S5. Growth curve experiments showing cell proliferation upon TGFβ/Activin signaling inhibition. (C and G) Cell cycle analysis by FACS measuring BrdU incorporation and 7AAD labeling at 48 h upon inhibition. The percentage of cells in the different phases of the cell cycle is represented (G0/G1, S, and G2/M phases). (D and H) Percentage of apoptotic cells measured by FACS analysis of cleaved caspase‐3 48 h after TGFβ/Activin signaling inhibition. The P‐values were determined by unpaired t‐test and two‐way ANOVA for (B and F). *P < 0.05, **P < 0.01, ***P < 0.001. Bars represent the mean ± SD. Number of replicates is n ≥ 3. The exact P‐values and number of replicates are indicated in Appendix Table S5.

Source data are available online for this figure.