Figure EV3. PMEPA1 is a target gene involved in the response to Activin signaling.

• A
  Immunoblots of phosphorylated Smad2 (P‐Smad2), total Smad2, MYC, PMEPA1, OTX2, and β‐actin were performed on cell extracts from 1603MED transfected with control siRNA control (siCTRL, blue) or targeting INHBB (siINHBB, red). 1603MED cells transfected with siINHBB were also treated with ActivinB to investigate a potential rescue (black). Lysates were prepared 48 h after transfection. Right panel represents WB quantification. The relative level to β‐actin is presented for the different proteins. Note that P‐Smad2/β‐actin quantification is identical to that of in Figs 2D and EV2A. The level in control conditions (siCTRL) was set at 1.
• B–E
  D283 (B and C) and 1603MED (D and E) cell lines were transfected with siCTRL (blue) or individual siRNAs targeting PMEPA1 (siPMEPA1#1 or siPMEPA1#2 red). D283 (B) and 1603MED (D) lysates were prepared 48 h after transfection. Right panel represents WB quantifications performed as above. (C and E) Viable D283 (C) and 1603MED (E) cells measured 2 days after transfection with either siCTRL (blue) or siPMEPA1#1 and #2 (red).

Data information: The P‐values were determined by unpaired t‐test. *P < 0.05, **P < 0.01, and ***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.