Journal: Pharmaceuticals

Article Title: Glabridin Inhibits Melanogenesis and Melanin Transfer via Wnt/β-Catenin Pathway and Rho Family GTPase-Mediated Dendritic Formation Suppression

doi: 10.3390/ph19030469

Figure Lengend Snippet: Effect of glabridin on anti-melanogenesis. Different concentrations of glabridin on MNT-1 cell viability ( a ) and on HaCaT cell viability ( b ), assessed by the MTT assay; Different concentrations of niacinamide on MNT-1 cell viability ( c ) and on HaCaT cell viability ( d ), assessed by the MTT assay; ( e ) Glabridin inhibit melanin content in MNT-1 cells. ( f ) Glabridin inhibit intracellular tyrosinase activity in MNT-1 cells. The images of protein bands ( g ) and the relative protein levels of MITF ( h ), TYR ( i ), TRP-1 ( j ) and TRP-2 ( k ). GAPDH served as the internal control. Kojic acid (KA, 200 μg/mL) was used as a positive control. Data are presented as mean ± SD ( n = 3) (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. control) (one-way ANOVA followed by Tukey’s post hoc test).

Article Snippet: Briefly, membranes were blocked using QuickBlockTM Blocking Buffer for 15 min, after which they were incubated overnight (12 h) at 4 °C with antibodies against MITF (ET1702-86, HuaBio, Hangzhou, China), TYR (sc-20035, Santa Cruz, CA, USA), TRP-1 (sc-166857, Santa Cruz, CA, USA), TRP-2 (sc-74439, Santa Cruz, CA, USA), Rac1 (sc-514583, Santa Cruz, CA, USA), RhoA (sc-418, Santa Cruz, CA, USA), Cdc42 (ET1701-7, HuaBio, Hangzhou, China) and GAPDH (60004-1-lg, Proteintech, Wuhan, China).

Techniques: MTT Assay, Activity Assay, Control, Positive Control

Journal: Pharmaceuticals

Article Title: Glabridin Inhibits Melanogenesis and Melanin Transfer via Wnt/β-Catenin Pathway and Rho Family GTPase-Mediated Dendritic Formation Suppression

doi: 10.3390/ph19030469

Figure Lengend Snippet: Glabridin inhibits UVB-induced melanin transfer in MNT-1/HaCaT co-culture systems. ( a ) Masson-Fontana staining of the co-culture system. Red arrows: UVB-induced melanocytes with enhanced melanogenesis and elongated dendrites. Green arrows: glabridin-treated melanocytes showing reduced melanin content and shortened dendrites. Yellow boxes: melanosome transferred to HaCaT cells. Niacinamide (NAM, 200 μM) served as a positive control. Scale bar, 100 μm. ( b , c ) Melanin content in MNT-1 cells ( b ) and HaCaT cells ( c ) after UVB irradiation followed by treatment with niacinamide and different concentrations of glabridin. ( d ) Quantification of TRP-1/pan-cytokeratin co-localization coefficients. ( e ) Immunofluorescence staining showing TRP-1 (melanosome marker, red) and pan-cytokeratin (keratinocyte marker, green) localization. White boxes indicate regions of co-localization. Scale bar, 100 μm. Data are presented as mean ± SD ( n = 3) (** p < 0.01 vs. the untreated control group; # p < 0.05, ## p < 0.01, ### p < 0.001, and #### p < 0.0001 vs. UVB-treated group) (one-way ANOVA followed by Tukey’s post hoc test).

Article Snippet: Briefly, membranes were blocked using QuickBlockTM Blocking Buffer for 15 min, after which they were incubated overnight (12 h) at 4 °C with antibodies against MITF (ET1702-86, HuaBio, Hangzhou, China), TYR (sc-20035, Santa Cruz, CA, USA), TRP-1 (sc-166857, Santa Cruz, CA, USA), TRP-2 (sc-74439, Santa Cruz, CA, USA), Rac1 (sc-514583, Santa Cruz, CA, USA), RhoA (sc-418, Santa Cruz, CA, USA), Cdc42 (ET1701-7, HuaBio, Hangzhou, China) and GAPDH (60004-1-lg, Proteintech, Wuhan, China).

Techniques: Co-Culture Assay, Staining, Positive Control, Irradiation, Immunofluorescence, Marker, Control

Journal: Frontiers in oncology

Article Title: YTHDF1 Aggravates the Progression of Cervical Cancer Through m 6 A-Mediated Up-Regulation of RANBP2.

doi: 10.3389/fonc.2021.650383

Figure Lengend Snippet: FIGURE 5 | RANBP2 is the key target of YTHDF1 in cervical cancer. (A) Western blot detecting the protein level of RANBP2 in Hela and Siha cells upon YTHDF1 knockdown. (B) RT-qPCR detecting relative RNA level of RANBP2 in Hela and Siha upon YTHDF1 knockdown. (C) RIP-PCR assays detecting the interactions between YTHDF1 and RANBP2 mRNA in Siha cells. IgG was used as an internal control. GAPDH was used as the negative control in western blot assays. (D) meRIP-PCR assays detecting the m6A modification of RANBP2 mRNA in Siha cells. (E) Schematic of wild-type (YTHDF1-wt) and mutant (YTHDF1-mut) YTHDF1 constructs. (F) RIP-derived RNA and protein of wild-type (YTHDF1-wt) group and mutant (YTHDF1-mut) group in Hela cells were measured by RT-qPCR and western blot after immunoprecipitation by using the antibody specific to Flag, respectively. GAPDH was used as the negative control in western blot assays. Data are shown as means ± S.D. **P < 0.01, ***P < 0.001.

Article Snippet: The antibodies used for western blot are as follows: YTHDF1 (ProteinTech, 1:1000), RANBP2 (ProteinTech, 1:1000), GAPDH (ZSGB-BIO, 1:2000), Flag-tag (MBL, 1:5000).

Techniques: Western Blot, Knockdown, Quantitative RT-PCR, Control, Negative Control, Mutagenesis, Construct, Derivative Assay, Immunoprecipitation

Journal: Frontiers in oncology

Article Title: YTHDF1 Aggravates the Progression of Cervical Cancer Through m 6 A-Mediated Up-Regulation of RANBP2.

doi: 10.3389/fonc.2021.650383

Figure Lengend Snippet: FIGURE 6 | RANBP2 plays an oncogenic role in cervical cancer cells. (A) Detection of RANBP2 knockdown in Hela and Siha cell lines by western blot. (B) The effect of RANBP2 knockdown on cell growth was determined by CCK-8 assays. (C) Colony formation assays were performed in RANBP2 knockdown and control cells. (D, E) Migration and invasion assays of Hela and Siha cells upon RANBP2 knockdown. Scale bar, 200 mm. Data are shown as means ± S.D. **P < 0.01, ***P < 0.001.

Article Snippet: The antibodies used for western blot are as follows: YTHDF1 (ProteinTech, 1:1000), RANBP2 (ProteinTech, 1:1000), GAPDH (ZSGB-BIO, 1:2000), Flag-tag (MBL, 1:5000).

Techniques: Knockdown, Western Blot, CCK-8 Assay, Control, Migration

Journal: Frontiers in oncology

Article Title: YTHDF1 Aggravates the Progression of Cervical Cancer Through m 6 A-Mediated Up-Regulation of RANBP2.

doi: 10.3389/fonc.2021.650383

Figure Lengend Snippet: FIGURE 7 | Knockdown of RANBP2 suppressed the proliferation, migration and invasion of YTHDF1-overexpressing Hela and Siha cells. (A) Colony formation assays were performed in YTHDF1-overexpressing Hela and Siha cells infected with the RANBP2 shRNA or controls. (B) The proliferation ability of YTHDF1- overexpressing Hela and Siha cells upon RANBP2 knockdown was assessed by CCK-8 assays. (C, D) Migration and invasion YTHDF1-overexpressing Hela (C) and Siha (D) cells upon RANBP2 knockdown was detected by transwell assays. Scale bar, 200 mm. *P < 0.05,**P < 0.01, ***P < 0.001.

Article Snippet: The antibodies used for western blot are as follows: YTHDF1 (ProteinTech, 1:1000), RANBP2 (ProteinTech, 1:1000), GAPDH (ZSGB-BIO, 1:2000), Flag-tag (MBL, 1:5000).

Techniques: Knockdown, Migration, Infection, shRNA, CCK-8 Assay

Journal: Frontiers in oncology

Article Title: YTHDF1 Aggravates the Progression of Cervical Cancer Through m 6 A-Mediated Up-Regulation of RANBP2.

doi: 10.3389/fonc.2021.650383

Figure Lengend Snippet: FIGURE 8 | The expression of RANBP2 is positively correlated with YTHDF1 in cervical cancer. (A) Representative immunohistochemical images of RANBP2 protein expression in cervical cancer tissues and cervical epithelium tissues. Scale bar, 100 mm. (B) The quantitative analysis of RANBP2 expression in cervical cancer tissues and cervical epithelium tissues assessed by immunohistochemistry. (C) Spearman’s correlation analysis of RANBP2 and YTHDF1 expression in cervical cancer tissues. (D) Representative immunohistochemical images of YTHDF1 and RANBP2 in the cervical cancer tissues. Scale bar, 100 mm. Data are shown as means ± S.D. *P < 0.05.

Article Snippet: The antibodies used for western blot are as follows: YTHDF1 (ProteinTech, 1:1000), RANBP2 (ProteinTech, 1:1000), GAPDH (ZSGB-BIO, 1:2000), Flag-tag (MBL, 1:5000).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry