Journal: Journal of Crohn's & Colitis

Article Title: CD147 Targeting by AC-73 Induces Autophagy and Reduces Intestinal Fibrosis Associated with TNBS Chronic Colitis

doi: 10.1093/ecco-jcc/jjac084

Figure Lengend Snippet: AC-73 inhibits activation of the CD147/ERK1/2 and STAT3 signalling pathways and induces autophagy in mice with chronic TNBS colitis. [A, B] Western blot analysis of [A] ERK1/2 total [pan ERK1/2] and phosphorylated ERK1/2 [pERK1/2] protein expression levels, [B] STAT3 total [pan STAT3] and phosphorylated STAT3 [pSTAT3] protein expression levels. [A, B] Left: densitometric data are presented as the ratio of activated ERK to total ERK [pERK1/2/pan ERK1/2] and the ratio of activated STAT3 to total STAT3 [p-STAT3/pan STAT3]. [C] Western blot analysis of autophagy-related protein LC3 and its conversion from LC3-I to LC3-II in TNBS+AC-73, TNBS+V and ETOH+V mice; one representative western blot out of three is shown with densitometric data of the LC3B-II to LC3B-I ratio; GAPDH is an internal control of total protein extracts; right: densitometric data [A.U. = arbitrary units] represented as individual plots from at least: ten TNBS-treated mice per group and five ethanol-treated mice. Lines indicate median values. ns [not significant], * p <0.05, ** p <0.01 from a Mann–Whitney test.

Article Snippet: Antibodies used were: CD147 monoclonal antibody (EMMPRIN/CD147 [B5] sc-46700, Santa Cruz); TGF-β1 monoclonal antibody (TGFβ1[TB21] sc-52893, Santa Cruz); α-SMA polyclonal antibody [Cell Signaling Technology, Ma, USA]; for total STAT3 protein detection, STAT3 [H-190] polyclonal antibody [sc-7179, Santa Cruz]; phospho-STAT3-S727 and phospho-ERK1/2 [p44/p42 MAPK] polyclonal antibodies [Immunological Sciences]; for total ERK1/2 protein detection, p44/42 MAPK [Erk1/2] polyclonal antibody [9102, Cell Signaling Technology]; GAPDH polyclonal antibody [Sigma-Aldrich] was used as an internal control of the loaded amounts of total proteins.

Techniques: Activation Assay, Western Blot, Expressing, MANN-WHITNEY