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Aberrant activation of the Raf/ERK signaling pathway in BMPR2-silenced HPAECs. Lysates from control HPAECs and BMPR2-deficient HPAECs were resolved by SDS-PAGE and immunoblotted. pRAF1 (serine 338) and total RAF1 (3 independent experiments in 3 different donors) (A): pARAF (Ser-445) and total <t>ARAF</t> (n = 4) (B), pBRAF (Ser-445) and total BRAF (n = 4) (C), pERK, and total ERK (n = 6) (D), pp38 and total p38 (n = 4) (E), pJNK and total JNK (n = 4) (F), and BIM (BCL2L11; n = 4) (G). Densitometric analyses are normalized to the loading controls and the corresponding siControl condition. Representative blots are shown, and data are presented as means ± SE; *P < 0.05; **P < 0.01. H: schematic diagram of dysregulated Ras/Raf/ERK signaling resulting from BMPR2 loss-of-function. Red (up-regulated) and green (down-regulated) shading indicates the directional changes of significantly regulated transcripts [false discovery rates (FDR) < 0.01; no FC cutoff] by microarray. Black arrows (up or down) indicate experimentally determined directional changes in protein expression and/or activation state.
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Aberrant activation of the Raf/ERK signaling pathway in BMPR2-silenced HPAECs. Lysates from control HPAECs and BMPR2-deficient HPAECs were resolved by SDS-PAGE and immunoblotted. pRAF1 (serine 338) and total RAF1 (3 independent experiments in 3 different donors) (A): pARAF (Ser-445) and total <t>ARAF</t> (n = 4) (B), pBRAF (Ser-445) and total BRAF (n = 4) (C), pERK, and total ERK (n = 6) (D), pp38 and total p38 (n = 4) (E), pJNK and total JNK (n = 4) (F), and BIM (BCL2L11; n = 4) (G). Densitometric analyses are normalized to the loading controls and the corresponding siControl condition. Representative blots are shown, and data are presented as means ± SE; *P < 0.05; **P < 0.01. H: schematic diagram of dysregulated Ras/Raf/ERK signaling resulting from BMPR2 loss-of-function. Red (up-regulated) and green (down-regulated) shading indicates the directional changes of significantly regulated transcripts [false discovery rates (FDR) < 0.01; no FC cutoff] by microarray. Black arrows (up or down) indicate experimentally determined directional changes in protein expression and/or activation state.
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Aberrant activation of the Raf/ERK signaling pathway in BMPR2-silenced HPAECs. Lysates from control HPAECs and BMPR2-deficient HPAECs were resolved by SDS-PAGE and immunoblotted. pRAF1 (serine 338) and total RAF1 (3 independent experiments in 3 different donors) (A): pARAF (Ser-445) and total ARAF (n = 4) (B), pBRAF (Ser-445) and total BRAF (n = 4) (C), pERK, and total ERK (n = 6) (D), pp38 and total p38 (n = 4) (E), pJNK and total JNK (n = 4) (F), and BIM (BCL2L11; n = 4) (G). Densitometric analyses are normalized to the loading controls and the corresponding siControl condition. Representative blots are shown, and data are presented as means ± SE; *P < 0.05; **P < 0.01. H: schematic diagram of dysregulated Ras/Raf/ERK signaling resulting from BMPR2 loss-of-function. Red (up-regulated) and green (down-regulated) shading indicates the directional changes of significantly regulated transcripts [false discovery rates (FDR) < 0.01; no FC cutoff] by microarray. Black arrows (up or down) indicate experimentally determined directional changes in protein expression and/or activation state.

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Raf/ERK drives the proliferative and invasive phenotype of BMPR2-silenced pulmonary artery endothelial cells

doi: 10.1152/ajplung.00303.2015

Figure Lengend Snippet: Aberrant activation of the Raf/ERK signaling pathway in BMPR2-silenced HPAECs. Lysates from control HPAECs and BMPR2-deficient HPAECs were resolved by SDS-PAGE and immunoblotted. pRAF1 (serine 338) and total RAF1 (3 independent experiments in 3 different donors) (A): pARAF (Ser-445) and total ARAF (n = 4) (B), pBRAF (Ser-445) and total BRAF (n = 4) (C), pERK, and total ERK (n = 6) (D), pp38 and total p38 (n = 4) (E), pJNK and total JNK (n = 4) (F), and BIM (BCL2L11; n = 4) (G). Densitometric analyses are normalized to the loading controls and the corresponding siControl condition. Representative blots are shown, and data are presented as means ± SE; *P < 0.05; **P < 0.01. H: schematic diagram of dysregulated Ras/Raf/ERK signaling resulting from BMPR2 loss-of-function. Red (up-regulated) and green (down-regulated) shading indicates the directional changes of significantly regulated transcripts [false discovery rates (FDR) < 0.01; no FC cutoff] by microarray. Black arrows (up or down) indicate experimentally determined directional changes in protein expression and/or activation state.

Article Snippet: Antibodies against p-ERK (no. 9101, 1:1,000), total ERK (no. 9102, 1:1,000), p-p38 (no. 9211, 1:400), total p38 (no. 9212, 1:800), p-JNK (no. 4671, 1:400), total JNK (no. 9258, 1:1,000), p-RAF-1 (no. 9427, 1:1,000), p-ARAF (no. 4431, 1:500), p-BRAF (no. 2696, 1:500), were purchased from Cell Signaling (Danvers, MA).

Techniques: Activation Assay, Control, SDS Page, Microarray, Expressing