Anti msk1

Immunoblotting was carried out as described previously [11 (link)]. The following antibodies were purchased from Abcam: anti-GAPDH (1:2500, 37 kDa, AB9485, RRID:AB_307275), anti-IL-6 (1:2000, 17 and 50 kDa, ab9324, RRID:AB_307175), and anti-ICAM-1 (1:4000, 90 kDa, AB53013, RRID:AB_870702). Anti-ACE2 (1:1000, 125 kDa, MA532307, RRID:AB_2809589) and anti-HIF-1α (1:1000, 130 kDa, PA5-85,494, RRID:AB_2792634) antibodies were obtained from Thermo Scientific. The following antibodies were purchased from Cell Signaling Technology: anti-ERK1/2 (1:1000, 42, 44 kDa, #4695, RRID:AB_390779), anti-phospho-ERK1/2 (Thr202/Tyr204), (1:1000, 42, 44 kDa, # 9101, RRID:AB_331646), anti-MSK1 (1:1000, 95 kDa, #3489, RRID:AB_2285349), anti-phospho-MSK1 (Thr581) (1:1000, 95 kDa, #9595, RRID:AB_2181783), anti-p38 (1:1000, 43 kDa, #9212, RRID:AB_330713), anti-phospho-p38 (Thr180/Tyr182) (1:1000, 43 kDa, #4511, RRID:AB_2139682), anti-phospho-JNK (Thr183/Tyr185) (1:1000, 46, 54 kDa, #4668, RRID:AB_823588), anti-JNK (1:1000, 46, 64 kDa, #9252, RRID:AB_2250373), anti-phospho-NFκB p65 (Ser536) (1:1000, 70 kDa, #3033, RRID:AB_331284), and anti-NFκB p65. (1:1000, 70 kDa, #6956, RRID:AB_10828935). The following antibody were obtained from ABclonal (Woburn, MA): anti-phospho-NFkB p65 (S276) (1:1000, 70 kDa, AP0123, RRID:AB_2771505). An anti-phospho-MBP antibody (1:200, 18 kDa, 13–104) was purchased from Merck.

Dexamethasone (D4902) (DEX) and GW7647 (G6793) (GW) were obtained from Sigma–Aldrich (St. Louis, MO, USA). Anti-GR, anti-PPARα, anti-RNA pol II and anti-p65 antibodies were obtained from Santa Cruz. Phospho-specific rabbit antibodies to p38 (Thr-180/Tyr-182), p42/44 ERK (Thr202/Tyr204), MSK1 (Thr581) and IKKα/β (Ser180/S181) were used to detect the respective phosphorylated forms and purchased from Cell Signaling. Anti-p38, anti-ERK, anti-MSK1, and anti-IκBα antibodies were purchased from Cell Signaling. Anti-tubulin and anti-actin were used as loading control and obtained from Santa Cruz. Anti-phospho-65 was obtained from Santa Cruz. Recombinant murine TNFα was produced and purified as described (30 (link)). TNFα was used at a final concentration of 2,000 IU/ml. p(IL6κB)₃50hu.IL6P-luc+ (hereafter renamed NF-κB-Luc), PPARα, GR, and 5HT7 control plasmids were described previously (21 (link), 31 (link)–33 (link)). LPS was purchased from Invivogen.

An anti‐FLAG antibody (M2) was purchased from Merck Millipore (St. Charles, MO, USA). Anti‐cyclin A (SC‐751), anti‐cyclin D1 (SC‐753), anti‐CDC2 (SC‐163), anti‐CDK4 (SC‐601), anti‐H‐Ras (SC‐520), and anti‐β‐actin (SC‐130301) antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). An anti‐phospho p65/RelA (S276) antibody was purchased from Rockland Immunochemicals (Limerick, PA, USA). Anti‐MSK1 and anti‐MSK2 antibodies were purchased from Cell Signaling Technology (#9309; Danvers, MA, USA) and BD Biosciences (San Jose, CA, USA), respectively. TNFα and PDGF were purchased from PeproTech (Rocky Hill, NJ, USA). LY294002, U0126, SP600125, and SB203580 were obtained from FUJIFILM Wako Pure Chemical (Osaka, Japan). NOX inhibitor, VAS2870, was purchased from Merck Millipore (Burlington, MA, USA).