Journal: International Journal of Molecular Sciences

Article Title: Chemerin-156 is the Active Isoform in Human Hepatic Stellate Cells

doi: 10.3390/ijms21207555

Figure Lengend Snippet: Alpha-smooth muscle actin (α-SMA) and galectin-3 in LX-2 cells expressing chemerin isoforms. ( a – c ) LX-2 cells were transfected with an insertless plasmid (C) and plasmids to express huChem-155, 156 and 157. Cellular α-SMA was analyzed by immunoblot at 24, 48 and 72 h post-transfection. Coomassie stained membrane served as control; ( d – f ) Galectin-3 in the cells described in a–c; ( g – i ) Galectin-3 in the cell media of LX-2 cells described in a–c. N = 3; * p < 0.05, ** p < 0.01, *** p < 0.001, the number in ( h ) in brackets is the p -value.

Article Snippet: The antibodies used were obtained from the following sources: human chemerin, R&D Systems (Wiesbaden-Nordenstadt, Germany); GAPDH, New England Biolabs GmbH (Frankfurt am Main, Germany); galectin-3, BD Biosciences (Heidelberg, Germany); α-SMA and CMKLR1, Abcam (Cambridge, UK); GPR1, Bioss (Woburn, MA, USA) and Abnova (Taipei City, Taiwan).

Techniques: Expressing, Transfection, Plasmid Preparation, Western Blot, Staining

Journal: International Journal of Molecular Sciences

Article Title: Chemerin-156 is the Active Isoform in Human Hepatic Stellate Cells

doi: 10.3390/ijms21207555

Figure Lengend Snippet: Secretome of HSCs and LX-2 cells. ( a ) IL-6 in the supernatant of HSCs of three different donors and LX-2 cells; ( b ) Pentraxin 3 (PTX3); ( c ) IL-8; ( d ) galectin-3; and ( e ) chemerin in the cell culture media of these cells.

Article Snippet: The antibodies used were obtained from the following sources: human chemerin, R&D Systems (Wiesbaden-Nordenstadt, Germany); GAPDH, New England Biolabs GmbH (Frankfurt am Main, Germany); galectin-3, BD Biosciences (Heidelberg, Germany); α-SMA and CMKLR1, Abcam (Cambridge, UK); GPR1, Bioss (Woburn, MA, USA) and Abnova (Taipei City, Taiwan).

Techniques: Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Chemerin-156 is the Active Isoform in Human Hepatic Stellate Cells

doi: 10.3390/ijms21207555

Figure Lengend Snippet: Effect of huChem-157 on IL-6, IL-8, pentraxin 3 (PTX3) and galectin-3 in primary human HSCs of three donors. ( a ) HSCs were incubated with increasing concentrations of recombinant huChem-157 for 24 h and IL-6; ( b ) IL-8; ( c ) PTX3 and ( d ) galectin-3 were measured by ELISA.

Article Snippet: The antibodies used were obtained from the following sources: human chemerin, R&D Systems (Wiesbaden-Nordenstadt, Germany); GAPDH, New England Biolabs GmbH (Frankfurt am Main, Germany); galectin-3, BD Biosciences (Heidelberg, Germany); α-SMA and CMKLR1, Abcam (Cambridge, UK); GPR1, Bioss (Woburn, MA, USA) and Abnova (Taipei City, Taiwan).

Techniques: Incubation, Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Autophagy

Article Title: Periplocin suppresses the growth of colorectal cancer cells by triggering LGALS3 (galectin 3)-mediated lysophagy.

doi: 10.1080/15548627.2023.2239042

Figure Lengend Snippet: Figure 6. Periplocin binds and prevents ubiquitin-mediated degradation of LGALS3 in CRC cells. (A) Immunoblotting analysis of LGALS3 in cells treated with periplocin for 24 h at the indicated concentrations. (B and C) Representative images (B) and quantitative analysis (C) of immunohistochemical staining for LGALS3 in SW480 ×enografts from vehicle- or periplocin-treated mice. Scale bar: 50 μm. (D) Immunoblotting analysis of LGALS3 in cells treated with 0.50 μM periplocin for 24 h in the presence or absence of cycloheximide (CHX, 50 μg/mL). (E) Quantitation of LGALS3 protein level in (D). (F) Immunoblotting analysis of LGALS3 in cells treated with 0.50 μM periplocin for 24 h in the presence or absence of MG132 (25 μM, 6 h). (G) Quantitation of LGALS3 protein level in (F). (H) FLAG-LGALS3 was co-expressed

Article Snippet: LGALS3 (MAB1154) was purchased from R&D Systems.

Techniques: Ubiquitin Proteomics, Western Blot, Immunohistochemical staining, Staining, Quantitation Assay

Journal: Autophagy

Article Title: Periplocin suppresses the growth of colorectal cancer cells by triggering LGALS3 (galectin 3)-mediated lysophagy.

doi: 10.1080/15548627.2023.2239042

Figure Lengend Snippet: Figure 7. Periplocin induces lethal lysophagy by upregulating LGALS3 in CRC cells. (A and B) Immunoblotting analysis of LC3B turnover in cells transfected with siNC or siLGALS3 followed by 0.50 μM periplocin treatment for 24 h. (C and D) Immunoblotting analysis of LC3B turnover in parental or lgals3 KO cells followed by 0.50 μM periplocin treatment for 24 h. (E and F) Representative images (E) and quantitative analysis (F) for immunofluorescent staining of endogenous LC3B puncta in cells transfected with siNC or siLGALS3 followed by 0.50 μM periplocin treatment for 24 h. Scale bar: 10 μm. (G and H) MTT assay of CRC cells with or without LGALS3 knockout (G, lgals3 KO#1; H, lgals3 KO#2) in response to 0.50 μM periplocin treatment for 24 h. (I) Colony formation assay of parental or lgals3 KO cells treated with or without 0.50 μM periplocin for 24 h. (J) Quantification of clone numbers in (I). (K and L) DLD-1 parental or lgals3 KO cells were subcutaneously inoculated into nude mice. Mice were injected with vehicle or periplocin (15 mg/kg/day) for two weeks. Image (K) and weight (L) of tumor xenografts were shown. Results in A-J are representative of three independent experiments. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. ns, non-significant.

Article Snippet: LGALS3 (MAB1154) was purchased from R&D Systems.

Techniques: Western Blot, Transfection, Staining, MTT Assay, Knock-Out, Colony Assay, Injection