Journal: bioRxiv

Article Title: Cooperative phagocytosis underlies macrophage immunotherapy of solid tumours and initiates a broad anti-tumour IgG response

doi: 10.1101/2022.01.01.474150

Figure Lengend Snippet: Growth curves of mice bearing CD47 KO B16 tumours (2×10 5 s.c. flank injection) and treated i.v. on days 4, 5, 7, 9, 11, 13, and 15 with 250 µg isotype IgG2a control (clone C1.18.4), 100 µl PBS vehicle, or left untreated. Clone C1.18.4 is the isotype control antibody for anti-Tyrp1 clone TA99. There are no significant growth differences (mean ± SEM, n = 6 mice per group).

Article Snippet: Antibodies used for in vivo treatment and blocking, in vitro phagocytosis, flow cytometry, and Western blotting are as follows: anti-mouse/human Tyrp1 clone TA99 (BioXCell BE0151), isotype IgG2a control clone C1.18.4 (BioXCell, BE0085), anti-mouse CD47 clone MIAP301 (BioXCell BE0270), isotype control rat IgG2a (BioXCell, BE0089), anti-mouse SIRPα clone P84 (BioLegend 144004), isotype control rat IgG1 (BioLegend, 400414).

Techniques: Injection, 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: 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: Cell Death Discovery

Article Title: Lysine attenuates acute lung injury by restoring α-tubulin acetylation and ciliary activity

doi: 10.1038/s41420-026-03025-x

Figure Lengend Snippet: Calcium image analysis in PQ-injured A549 cells compared to normal cells ( A ) and PQ-injured A549 cells w/wo lysine supplementation ( B ) with thapsigargin (TG) or ionomycin (IONO) stimulation. Mean ± SEM. C NFAT luciferase expression in A549 cells treated w/wo 800 μM PQ, together with 0-, 1-, 2.5-, or 5-fold lysine as in the culture medium as indicated, for 24 h. Mean ± SD, *** P < 0.001; Two-Way ANOVA. n = 3. NS not significant. D WB analysis of E-Cadherin, ZO-1, EPCAM and acetyl-α-Tubulin in PQ-injured A549 cells treated w/wo 5 μM SKF-96365 (SKF). E Co-IP analysis of STIM1 association with ORAI1 or TRPC1 in PQ-injured A549 cells with indicated concentration, together w/wo 5 mM lysine. F Immunofluorescence analysis of Myc and ARL13B in PQ-injured TRPC1-Myc stably expressed A549 cells w/wo lysine supplementation. Scale bar, 10 μm.

Article Snippet: Specific antibodies were used for STIM1 (Cell Signaling Technology, 5668, 1:3000), ORAI1 (Santa Cruz, sc-377281, 1:500), and TRPC1 (Proteintech, 19482, 1:2000).

Techniques: Luciferase, Expressing, Co-Immunoprecipitation Assay, Concentration Assay, Immunofluorescence, Stable Transfection

Journal: Cell Death Discovery

Article Title: Lysine attenuates acute lung injury by restoring α-tubulin acetylation and ciliary activity

doi: 10.1038/s41420-026-03025-x

Figure Lengend Snippet: A GSEA analysis uncovered top-ranked molecular functions enhanced in PQ-injured lung tissues with lysine supplementation compared to the untreated group. B KEGG enrichment of the top 20 upregulated pathways in PQ-injured lung tissues with lysine supplementation compared to the untreated group. C Quantification of NADP and NADPH (left) or NAD and NADH (right) in PQ-injured A549 cells w/wo lysine supplementation. D ELISA analysis of acetyl coenzyme A (Acetyl-CoA) in PQ-injured A549 cells w/wo lysine supplementation. E WB analysis of E-Cadherin and acetyl-α-Tubulin in PQ-injured A549 cells treated w/wo 1 mM sodium acetate (SA). F WB analysis of ZO-1, E-Cadherin, acetyl-α-Tubulin and EPCAM in PQ-injured A549 cells w/wo lysine supplementation, together w/wo 0.5 μM GM-90257 (GM). G Calcium image analysis in PQ-injured A549 cells w/wo lysine supplementation, together w/wo 0.5 μM GM-90257 treatment. The cells were transiently stimulated with thapsigargin (TG) or ionomycin (IONO). Mean ± SEM. H Co-IP analysis of STIM1 association with TRPC1 in PQ-injured A549 cells w/wo lysine supplementation, together w/wo GM-90257 treatment. Immunofluorescence analysis of ARL13B, SFTPC, and HOPX in A549 cells ( I ) or normal lung tissues ( J ). Scale bar, 10 μm. Mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001; Two-Way ANOVA. NS not significant.

Article Snippet: Specific antibodies were used for STIM1 (Cell Signaling Technology, 5668, 1:3000), ORAI1 (Santa Cruz, sc-377281, 1:500), and TRPC1 (Proteintech, 19482, 1:2000).

Techniques: Enzyme-linked Immunosorbent Assay, Co-Immunoprecipitation Assay, Immunofluorescence