Journal: BMC Pulmonary Medicine

Article Title: Salt-inducible kinase 2 (SIK2) inhibitor ARN-3236 attenuates bleomycin-induced pulmonary fibrosis in mice

doi: 10.1186/s12890-022-01940-0

Figure Lengend Snippet: Inactivation of SIK2 led to the dephosphorylation and nuclear translocation of CRTC2 in human fetal lung fibroblasts (HFLs). HFLs or SIK2 overexpressed HFLs (HFLs-SIK2) were treated with DMSO (0.1%), ARN-3236 (0.5 μM) or transfected with SIK2 SiRNA (100 nM). Cells were then treated with TGF-β1 (5 ng/mL) for 12 h. A , B Representative western blot bands and quantitation of SIK2, pCRTC1, pCRTC2 and CRTC2 protein expressions in HFLs. C Representative confocal images and quantification of the nuclear translocated CRTC2 in HFLs and HFLs-SIK2 cells. Data are expressed as mean ± SEM of at least 4 independent experiments. *** P < 0.001; ** P < 0.01; * P < 0.05

Article Snippet: SIK2 inhibitor ARN-3236 was purchased from Medchemexpress (HY-120856) and Tsbiochem (T5993). pCREB inhibitor 666-15 was purchased from Medchemexpress (HY-101120) [ ], Tsbiochem (T5318) and Cayman Chem (30780).

Techniques: De-Phosphorylation Assay, Translocation Assay, Transfection, Western Blot, Quantitation Assay

Journal: BMC Pulmonary Medicine

Article Title: Salt-inducible kinase 2 (SIK2) inhibitor ARN-3236 attenuates bleomycin-induced pulmonary fibrosis in mice

doi: 10.1186/s12890-022-01940-0

Figure Lengend Snippet: Inactivation of SIK2 inhibited the fibroblast differentiation and TGF-β1 induced pro-fibrotic mediator expression in HFLs. HFLs or SIK2 overexpressed HFLs (HFLs-SIK2) were treated with DMSO (0.1%), ARN-3236 (0.5 μM), 666-15 (0.5 μM) or transfected with SIK2 SiRNA (100 nM). Cells were then treated with TGF-β1 (5 ng/mL) for 12 h. Inactivation of SIK2 inhibited A mRNA expression of α-SMA, COL1A and fibronectin and B cell proliferation in HFLs. Data are expressed as mean ± SEM of at least 4 independent experiments. *** P < 0.001; ** P < 0.01; * P < 0.05

Article Snippet: SIK2 inhibitor ARN-3236 was purchased from Medchemexpress (HY-120856) and Tsbiochem (T5993). pCREB inhibitor 666-15 was purchased from Medchemexpress (HY-101120) [ ], Tsbiochem (T5318) and Cayman Chem (30780).

Techniques: Expressing, Transfection

Journal: BMC Pulmonary Medicine

Article Title: Salt-inducible kinase 2 (SIK2) inhibitor ARN-3236 attenuates bleomycin-induced pulmonary fibrosis in mice

doi: 10.1186/s12890-022-01940-0

Figure Lengend Snippet: Inhibition of SIK2 reduced CRTC2 phosphorylation in mice. Male BALB/C mice were anesthetized and received intranasal instillation of BLM (5 mg/kg) or its saline vehicle, followed by receiving administration of ARN-3236 (10, 30 mg/kg) or its vehicle. A Representative western blot bands and quantitation of SIK2 and pCRTC2 protein expressions in lungs at day 14 and 28. B Representative western blot bands and quantitation of pCRTC2 protein expressions in lungs at day 28. Data are expressed as mean ± SEM of at least 6 independent experiments. *** P < 0.001; ** P < 0.01; * P < 0.05

Article Snippet: SIK2 inhibitor ARN-3236 was purchased from Medchemexpress (HY-120856) and Tsbiochem (T5993). pCREB inhibitor 666-15 was purchased from Medchemexpress (HY-101120) [ ], Tsbiochem (T5318) and Cayman Chem (30780).

Techniques: Inhibition, Saline, Western Blot, Quantitation Assay

Journal: BMC Pulmonary Medicine

Article Title: Salt-inducible kinase 2 (SIK2) inhibitor ARN-3236 attenuates bleomycin-induced pulmonary fibrosis in mice

doi: 10.1186/s12890-022-01940-0

Figure Lengend Snippet: Inhibition of SIK2 attenuates bleomycin (BLM)-induced pulmonary fibrosis in mice. Male BALB/C mice were anesthetized and received intranasal instillation of BLM (5 mg/kg) or its saline vehicle, followed by receiving administration of ARN-3236 (10, 30 mg/kg), CREB inhibitor 666-15 (10 mg/kg) or their vehicle. Lung tissues were isolated at day 28 for analysis. A Lung coefficient measured at day 28. B Mice body weight and lung coefficient measured at day 0, 14 and 28. C The content of hydroxyproline in lung tissues. D Semiquantitative analysis of lung tissues by H&E staining. E Representative histopathological sections of lung tissues by H&E staining. Data are expressed as mean ± SEM of at least 6 independent experiments. *** P < 0.001; ** P < 0.01; * P < 0.05

Article Snippet: SIK2 inhibitor ARN-3236 was purchased from Medchemexpress (HY-120856) and Tsbiochem (T5993). pCREB inhibitor 666-15 was purchased from Medchemexpress (HY-101120) [ ], Tsbiochem (T5318) and Cayman Chem (30780).

Techniques: Inhibition, Saline, Isolation, Staining

Journal: BMC Pulmonary Medicine

Article Title: Salt-inducible kinase 2 (SIK2) inhibitor ARN-3236 attenuates bleomycin-induced pulmonary fibrosis in mice

doi: 10.1186/s12890-022-01940-0

Figure Lengend Snippet: Inhibition of SIK2 attenuates bleomycin (BLM)-induced pulmonary fibrosis in mice. Male BALB/C mice were anesthetized and received intranasal instillation of BLM (5 mg/kg) or its saline vehicle, followed by receiving administration of ARN-3236 (10, 30 mg/kg), CREB inhibitor 666-15 (10 mg/kg) or their vehicle. Lung tissues were isolated at day 28 for analysis. Semiquantitative analysis of A Masson’s trichrome staining in E , B α-SMA expression in F and C COL1A expression in G were scored. D mRNA expression of α-SMA, COL1A, TGF-β1 and fibronectin in lungs. E Representative histopathological sections and semiquantitative analysis of lung tissues by Masson’s trichrome staining. Representative images of the immunohistochemical F α-SMA and G COL1A expression in lung tissues. Data are expressed as mean ± SEM of at least 6 independent experiments. *** P < 0.001; ** P < 0.01; * P < 0.05

Article Snippet: SIK2 inhibitor ARN-3236 was purchased from Medchemexpress (HY-120856) and Tsbiochem (T5993). pCREB inhibitor 666-15 was purchased from Medchemexpress (HY-101120) [ ], Tsbiochem (T5318) and Cayman Chem (30780).

Techniques: Inhibition, Saline, Isolation, Staining, Expressing, Immunohistochemical staining