Protein Expression Analysis of Fibrosis Markers

The protein levels of α-SMA, COL1A1, Smad2, p-Smad2, PED7A, PED4A, PKA, p-PKA, total Ras-proximate-1 (Rap1), Guanosine-5′-triphosphate (GTP)-Rap1, and exchange protein directly activated by cAMP 1 (Epac1), cAMP-response element-binding protein (CREB) and p-CREB were examined by immunoblotting following the methods described before (Liu et al., 2018 (link)) using the antibodies listed below: anti-α-SMA (ab5694, Abcam), anti-COL1A1 (ab34710, Abcam), anti-Smad2 (ab40855, Abcam), anti-p-Smad2 (ab53100, Abcam), anti-PDE4A (ab14607, Abcam), anti-PDE4B (ab170939, Abcam), anti-PDE4C (ab170939, Abcam), anti-PDE4D (ab171750, Abcam), anti-PKA (BS-0520R, Woburn, MA, United States), p-PKA (ab75991, Abcam), anti-GTP-Rap1 (ab32373, Abcam), anti-Rap1 (ab14404, Abcam), anti-Epac1 (ab109415, Abcam), anti-CREB (ab31387, Abcam), anti-p-CREB (ab32096, Abcam), and anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (ab8245, Abcam) and then with HRP-conjugated secondary antibody. Enhanced chemilumescent (ECL) substrates (Millipore, MA, United States) were used for signals visualization using GAPDH as an endogenous protein for normalization.

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Zhang B., Gao L., Shao C., Deng M, & Chen L. (2021). Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway. Frontiers in Pharmacology, 12, 722040.

Publication 2021

ab5694 ab34710 ab40855 ab53100 ab14607 ab170939 ab171750 ab75991 ab32373 ab14404 ab109415 ab31387 ab32096 ab8245 ECL) substrates

Antibodies Antibody Camp response element binding protein Glyceraldehyde 3 phosphate dehydrogenase Guanosine triphosphate Pde4b Pde4d Protein Smad2

Corresponding Organization : Central South University

Other organizations : Jinan Stomatological Hospital

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Variable analysis

independent variables

Unspecified

dependent variables

Protein levels of α-SMA
Protein levels of COL1A1
Protein levels of Smad2
Protein levels of p-Smad2
Protein levels of PDE4A
Protein levels of PDE4B
Protein levels of PDE4C
Protein levels of PDE4D
Protein levels of PKA
Protein levels of p-PKA
Protein levels of total Ras-proximate-1 (Rap1)
Protein levels of Guanosine-5′-triphosphate (GTP)-Rap1
Protein levels of exchange protein directly activated by cAMP 1 (Epac1)
Protein levels of cAMP-response element-binding protein (CREB)
Protein levels of p-CREB

control variables

GAPDH (used as an endogenous protein for normalization)

positive controls

Unspecified

negative controls

Unspecified

Annotations

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