Journal: Materials & Design

Article Title: Multifunctional nanofiber-based dressings in coordination with adipose-derived stem cells for accelerated burn wound healing

doi: 10.1016/j.matdes.2025.113929

Figure Lengend Snippet: Fig. 4. (A) Detection of ADSCs proliferation by ADM coating at different times. Quantification of (B) EGF and (C) FGF2 in ADSCs paracrine products. *** P < 0.001 representing a significant difference as compared with ADM (3 min) and TLWDA.

Article Snippet: Human EGF enzyme-linked immunosorbent assay (ELISA) kit and human FGF2 ELISA kit were purchased from BOSTER (Wuhan, China).

Techniques:

Journal: Nature Methods

Article Title: A polarized FGF8 source specifies frontotemporal signatures in spatially oriented cell populations of cortical assembloids

doi: 10.1038/s41592-024-02412-5

Figure Lengend Snippet: a , Schematic summarizing factors and gene markers involved in human cortical area specification. b , Overview of the protocol used to treat neural organoids with patterning factors from 3 to 5 div. Diff, differentiation medium ± vitamin A. c , Percentage of organoids with detectable SP8 > GFP expression after treatment with rostralizing factors (left) or EMX1>mNeonGreen expression after treatment with caudalizing factors (right). CHRDL1, chordin like 1; FST, follistatin; SB, SB-431542 dual SMAD inhibitor; CER1, cerberus 1; CHIR, GSK-3β inhibitor CHIR99021; WNT1, Wnt family member 1. Data indicate the mean ± s.d. from three lines per treatment ( n = 15 organoids per condition and line; except n = 16 for FGF8 high and CHIR low; n = 20, n = 19 and n = 17 for Chir high, and n = 18, n = 19 and n = 17 for Wnt1 treatment for the three lines). P values from one-way analysis of variance (ANOVA; Tukey’s multiple-comparisons test) for comparisons to untreated conditions are provided. d , Experimental procedure for assembloid generation with the SP8 > GFP line and OrEBs. D, day after EB formation; Diff, differentiation medium ± vitamin A; ULA, ultra-low attachment plate . e , RT–qPCR analysis of FGF8 target genes expressed in the SP8 > GFP EBs severed from OrEB after 1 day of co-culture. Data are the log of expression over TBP , shown as the mean ± s.d. ( n = 6 EBs for 0%, n = 5 for 1%, n = 5 for 10% grown from three independent clones). P values are the results of one-way ANOVA. f , GFP intensity of the organoids from g . Whiskers are the minima to maxima, boxes represent the 25th to 75th percentiles (Q1 to Q3) and lines indicate the median ( n = 7 organoids for 0%, n = 11 for 1%, n = 19 for 10%, grown from three independent clones). P values are the results of one-way ANOVA. g , Images of organoids generated with the SP8 > GFP transgenic line and co-culture with OrEBs at 60 div (scale bars, 500 µm). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.1. a.u., arbitrary units.

Article Snippet: Levels of FGF8 protein in cell extracts and supernatants were measured using an ELISA kit (Cusabio CSB-E15861h) according to the manufacturer’s instruction.

Techniques: Expressing, Quantitative RT-PCR, Co-Culture Assay, Clone Assay, Generated, Transgenic Assay

Journal: Nature Methods

Article Title: A polarized FGF8 source specifies frontotemporal signatures in spatially oriented cell populations of cortical assembloids

doi: 10.1038/s41592-024-02412-5

Figure Lengend Snippet: a ) Schematic diagram of CAG > FGF8 reporter (top). Genotyping of the selected clones (bottom) shows the three heterozygotes selected. The PCR was designed to amplify a wild type AAVS1 (WT) amplicon, a left homology arm amplicon (LA), a right homology arm amplicon (RA), and an inner part (IP). b ) RT-qPCR analysis of pluripotency markers and differentiation gene T/BRA in the selected CAG > FGF8 expressing clones and parental wild-type cells. Values are mean ± SD (n = 4 biological replicates for wt; n = 3 biological replicates, one for each CAG > FGF8 clone). P-values resulting from one-way ANOVA (Tukey’s multiple comparisons test) among the different lines are: OCT4 wt vs. OCT4 CAG > FGF8, p = 0.9166; NANOG wt vs. NANOG CAG > FGF8, p = 0.9841; SOX2 wt vs. SOX2 CAG > FGF8, p = 0.9997; TBRA wt vs. TBRA CAG > FGF8, p > 0.9999. c ) Box plots showing RT-qPCR analysis of FGF8 and WNT1 genes in the selected clones of the CAG > FGF8 line compared to wt. Whiskers are min to max, boxes represent the 25th to 75th percentiles (Q1 to Q3); n = 6 from 3 independent clones. One-way ANOVA analysis is p < 0.0001 for FGF8 expression and p > 0.9999 for WNT1 expression. d ) Quantification of FGF8 protein levels in cell lysates (CELLS) and supernatants (SUP) measured by ELISA assay. Values are mean ± SD, n = 6 from 3 independent clones; P-values from two-sided unpaired t-tests are: CELLS wt vs CELLS CAG > FGF8, p = 0.0004; SUP wt vs SUP CAG > FGF8, p = 0.0823.

Article Snippet: Levels of FGF8 protein in cell extracts and supernatants were measured using an ELISA kit (Cusabio CSB-E15861h) according to the manufacturer’s instruction.

Techniques: Clone Assay, Amplification, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Nature Methods

Article Title: A polarized FGF8 source specifies frontotemporal signatures in spatially oriented cell populations of cortical assembloids

doi: 10.1038/s41592-024-02412-5

Figure Lengend Snippet: a ) Representative images of 4 div embryoid bodies co-cultured with OrEBs containing cells expressing FGF8 and tdTomato mixed at different percentage (0%, 1%, 10%), immunostained for the FGF8 downstream target ETV1 (green) (scale bars 50 µm, the SP8 > GFP organoid is outlined with a dashed contour). The experiment has been repeated 4 times with organoids derived from three independent clones. b ) Immunostaining of 60 div organoids generated with the SP8 > GFP transgenic line and co-cultured with OrEBs in Fig. , stained for the rostral marker LMO4 (scale bars 500 µm in the main panels, 50 µm for the insets). The SP8 > GFP organoid is outlined with a dashed contour. c ) Box plots showing the fraction of LMO4+ cells normalized to total DAPI+ cells in 0%, 1% and 10% FGF8 conditions at 60 div. Whiskers are min to max, boxes represent the 25th to 75th percentiles (Q1 to Q3); n = 8 organoids. P-values from one-way ANOVA (Tukey’s multiple comparisons tests) are: 0% vs 1%, p = 0.0019, 0% vs 10%, p < 0.0001, 1% vs 10%, p < 0.0001. d ) Immunostaining of organoids generated with the SP8 > GFP transgenic line and co-cultured with OrEBs as described in Fig. for 60 days, stained for the neural marker Sox1 and DAPI. Scale bars 500 µm. e ) Quantification of the number of Sox1+ cells over total (DAPI + ) cells. Data show mean ± SD (n = 2 assembloids for 0%, n = 4 assembloids for 1%, n = 3 assembloids for 10%). P-values for each comparison resulting from one-way ANOVA (Tukey’s multiple comparisons tests) are: 0% vs 1%, p = 0.113; 0% vs 10%, p = 0.2171; 1% vs 10%, p = 0.8893. f ) RT-qPCR analysis of anterior ( PAX6, SIX3 ) and optic cup ( SNAI2, PAX3, OTX2, LHX2, RAX ) markers from SP8 > GFP organoids severed from the co-culture experiment at 60 div. Data is Log of expression over TBP, shown as mean ± SD; n = 6 for 0% condition, n = 5 for 1% and 10% condition, from 3 independent clones. P-values from one-way ANOVA (Tukey’s multiple comparisons tests) are: RAX 0% vs. RAX 1%, p = 0.0718; RAX 0% vs. RAX10%, p = 0.0003; RAX 1% vs. RAX 10%, p = 0.0263; all the other comparisons are p < 0.0001. g ) GFP intensity per segment (P, proximal; M, medial; D, distal) in the 1% FGF8 condition from experiments shown in panel b . Each segmented line represents an individual organoid. n = 7 from 3 clones; ns, p = 0.5783 one-way ANOVA. Ns, non-significant. a.u., arbitrary units.

Article Snippet: Levels of FGF8 protein in cell extracts and supernatants were measured using an ELISA kit (Cusabio CSB-E15861h) according to the manufacturer’s instruction.

Techniques: Cell Culture, Expressing, Derivative Assay, Clone Assay, Immunostaining, Generated, Transgenic Assay, Staining, Marker, Comparison, Quantitative RT-PCR, Co-Culture Assay

Journal: Nature Methods

Article Title: A polarized FGF8 source specifies frontotemporal signatures in spatially oriented cell populations of cortical assembloids

doi: 10.1038/s41592-024-02412-5

Figure Lengend Snippet: a , Experimental procedure for elongated assembloids using mosaic OrEBs containing CAG>tdTOMATO and non-fluorescent CAG > FGF8 -expressing cells. Diff, differentiation medium ± vitamin A. b , Representative images of elongated cortical assembloids at 1 div in the PDMS molds (indicated by white arrows in (i); scale bar, 500 µm), at 7 div after removal from the molds and before Matrigel embedding (ii) or after embedding in large Matrigel droplets ((iii); scale bar, 5 mm), at 120 div in the six-well plate ((iv); scale bar, 5 mm). c , Position of the OrEB on elongated cortical assembloids length (as a percentage) at 15, 60 and 120 div. Values are the mean ± s.d. ( n = 9 organoids for 15 div, n = 11 for 60 div and 120 div grown from three independent clones). P values for comparisons among time points (one-way ANOVA Tukey’s multiple-comparisons test) are: 15 div versus 60 div, P = 0.9455; 15 div versus 120 div, P = 0.9781; 60 div versus 120 div, P > 0.9999. d , Images of elongated cortical assembloids generated with the SP8 > GFP transgenic line and mosaic OrEBs at 60 div (scale bars, 500 µm). Right, SP8 > GFP intensity per segment (P, M and D) in individual assembloids. Each segmented line represents an individual elongated cortical assembloid ( n > 2 from at least 2 clones). P values are the results of one-way ANOVA among segments per condition (0%, P = 0.9045; 1%, P < 0.0001 and 10%, P = 0.9828). e , f , Images of proximal and distal CPNE8 ( e ) or NR2F1 ( f ) stainings (scale bars, 50 µm). Bottom, fraction of CPNE8 + ( e ) or NR2F1 + cells ( f ) normalized to total (DAPI + ) cells in proximal and distal insets of controls (conCAs) and polCAs at 60 div. Whiskers are min to max, boxes represent the 25th to 75th percentiles (Q1 to Q3) and lines indicate the median; CPNE8: n = 20 insets for P and D from 3 conCAs, n = 42 insets for P and n = 39 for D from 3 polCAs; NR2F1: n = 60 insets for P and D from 6 conCAs, n = 40 insets for P and D from 4 polCAs. P values from one-way ANOVA (Tukey’s multiple-comparisons test) are: P = 0.9988 in CPNE8 proximal conCA versus distal conCAs, P = 0.8509 in NR2F1 proximal conCAs versus distal conCAs, P < 0.0001 for other comparisons; NS, not significant. g , g ′, Images of 60 div polCA immunostained with tdTomato in red, DAPI in blue and NR2F1 in white ( g ) or intensity rainbow ( g ′). Scale bar, 500 µm.

Article Snippet: Levels of FGF8 protein in cell extracts and supernatants were measured using an ELISA kit (Cusabio CSB-E15861h) according to the manufacturer’s instruction.

Techniques: Expressing, Clone Assay, Generated, Transgenic Assay

Journal: Nature Methods

Article Title: A polarized FGF8 source specifies frontotemporal signatures in spatially oriented cell populations of cortical assembloids

doi: 10.1038/s41592-024-02412-5

Figure Lengend Snippet: a-b ) Representative images of elongated assembloids (8 div) properly embedded maintaining elongated shape (a) compared to elongated assembloids from the same batch without matrigel, shrinking without spatial constrain (b). c ) Length of elongated assembloids after different days in vitro (div, div1 n = 8, div7 n = 6, div30 n = 4, div60 n = 24, div120 n = 22 assembloids grown from 3 independent clones). P-values resulting from one-way ANOVA Tukey’s multiple comparisons tests) among the different lines are: div1 vs div7, p = 0.0045; div1 vs div30, p = 0.9999; div7 vs div30, p = 0.0366; div7 vs div60, p = 0.9998; div7 vs div120, p = 0.51; div30 vs div60, p = 0.0049; div60 vs div120, p = 0.1825; div1 vs div60, div1 vs div120, div30 vs div120, p < 0.0001. d ) RT-qPCR analysis of neural markers (SOX2, SOX1, SP8 and LMO4), ventral marker NXK2.1 and differentiation markers SNAI1 and T-BRA, in elongated assembloids (eOrg) compared to round organoids (rOrg), after treatment with high FGF8 as described in Fig. . Values are mean ± SD, n = 3 in three lines (hESC H9, H1 and iPSC 178/5). P-values resulting from one-way ANOVA (Tukey’s multiple comparisons tests) are: eORGSOX2 d10 vs. rORGSOX2 d10, p = 0.0024; eORGSOX2 d30 vs. rORGSOX2 d30, p = 0.3144; eORGSOX2 d60 vs. rORGSOX2 d60, p = 0.9434; eORGSOX1 d10 vs. rORGSOX1 d10, p = 0.0009; eORGSOX1 d30 vs. rORGSOX1 d30, p = 0.1573; eORGSOX1 d60 vs. rORGSOX1 d60, p = 0.7266; eORGSP8 d10 vs. rORGSP8 d10, p = 0.0006; eORGSP8 d30 vs. rORGSP8 d30, p = 0.1891; eORGSP8 d60 vs. rORGSP8 d60, p = 0.8004; eORGLMO4 d10 vs. rORGLMO4 d10, p = 0.0114; eORGLMO4 d30 vs. rORGLMO4 d30, p = 0.8378; eORGLMO4 d60 vs. rORGLMO4 d60, p = 0.9821; eORGNKX2.1 d10 vs. rORGNKX2.1 d10, p = 0.0008; eORGNKX2.1 d30 vs. rORGNKX2.1 d30, p = 0.1345; eORGNKX2.1 d60 vs. rORGNKX2.1 d60, p = 0.9036; eORGSNAI1 d10 vs. rORGSNAI1 d10, p = 0.0026; eORGSNAI1 d30 vs. rORGSNAI1 d30, p = 0.6125; eORGSNAI1 d60 vs. rORGSNAI1 d60, p = 0.96; eORGT-BRA d10 vs. rORGT-BRA d10, p = 0.6815; eORGT-BRA d30 vs. rORGT-BRA d30, p = 0.3026; eORGT-BRA d60 vs. rORGT-BRA d60, p = 0.1297. e ) Fraction of TUNEL+ cells normalized to total DAPI+ cells in elongated versus conventional round organoids (div, days in vitro ). N = 36 insets for 4 div elongated, n = 33 for 4 div Round, n = 122 for 7 div elongated, n = 37 for 7 div Round, n = 81 for 30 div elongated, n = 82 for 30 div Round organoids from 3 lines per condition. P-values resulting from one-way ANOVA (Tukey’s multiple comparisons tests) are: 4 div eOrg vs. 4 div rOrg, p < 0.0001; 7 div eOrg vs. 7 div rOrg, p = 0.31; 30 div eOrg vs. 30 div rOrg, p = 0.0115. f ) Box plots showing the fraction of TUNEL+ cells normalized to total DAPI+ cells in proximal and distal insets of elongated assembloids (n = 18 insets for 4 div Proximal and Distal, n = 32 for 7 div Proximal, n = 33 for 7 div Distal, n = 40 for 30 div Proximal, n = 42 for 30 div Distal; organoids are grown from 3 lines per condition). P-values resulting from one-way ANOVA (Tukey’s multiple comparisons tests) are: 4 div proximal vs. 4 div distal, p = 0.6097; 7 div proximal vs. 7 div distal, p = 0.6559; 30 div proximal vs. 30 div distal, p = 0.9897. g ) Representative images of the OrEB localization throughout elongated assembloids growth quantified in Fig. (scale bars 500 µm). h ) Top, representative images of proximal and distal CTIP2 staining (scale bars 50 µm). Bottom, fraction of CTIP2+ cells normalized to total DAPI+ cells in proximal and distal insets of controls (conCA) and polCAs at 60 div (n = 50 insets for P and D conCAs, n = 40 insets for P and D polCAs from 3 organoids). P-values resulting from one-way ANOVA (Tukey’s multiple comparisons tests) are: proximal conCA vs. distal conCA, p = 0.8204; proximal polCA vs. distal polCA, p < 0.0001. i ) Top, representative images of proximal and distal LMO4 staining (scale bars 50 µm). Bottom, fraction of LMO4+ cells normalized to total DAPI+ cells in proximal and distal insets of control(tdt) and polCAs at 60 div (n = 61 insets for P and D from 6 conCAs, n = 51 insets for P and D from 4 polCA). P-values resulting from one-way ANOVA (Tukey’s multiple comparisons tests) are: proximal conCA vs. distal conCA, p > 0.9999; proximal polCA vs. distal polCA, p < 0.0001. ns, non-significant. All whiskers are min to max, boxes represent the 25th to 75th percentiles (Q1 to Q3) and lines indicate median. j-j’ ) Images of immunostained longitudinal sections of elongated assembloids at 60 div (scale bars 500 µm).

Article Snippet: Levels of FGF8 protein in cell extracts and supernatants were measured using an ELISA kit (Cusabio CSB-E15861h) according to the manufacturer’s instruction.

Techniques: In Vitro, Clone Assay, Quantitative RT-PCR, Marker, TUNEL Assay, Staining, Control

Journal: Nature Methods

Article Title: A polarized FGF8 source specifies frontotemporal signatures in spatially oriented cell populations of cortical assembloids

doi: 10.1038/s41592-024-02412-5

Figure Lengend Snippet: a ) UMAP embedding for the scRNA-seq dataset containing cells derived from dissections of three segments of polCA and control counterparts annotated by stress levels based on Gruffi analysis. b ) Sankey plot showing similar mapping of cells from individual segments of both control and polCA to stressed cells according to Gruffi’s results. c ) Box plots showing the number of cells expressing FGF8 normalized by the total number of cells per segment for the three replicates. The box displays the median, the inter-quartile range, the minimum and maximum values for each segment. d ) Dot plot for the top five markers for each cluster, genes with logFC> 3 are shown. LogFCs colors are scaled by gene. e ) UMAP embedding plots colored by expression levels of markers of radial glia ( NES, SOX2, VIM ), cycling radial glia ( TOP2A, MKI67 ), Cajal-Retzious cells ( RELN ), neurons ( DCX, TUBB3 ), interneuron progenitor cells ( DLX6-AS1, GAD2 ), LGE-derived progenitors ( GSX2 ), MGE-derived progenitors ( NKX2.1 ), retinal progenitor cells ( RORB, VSX2 ), stress responsive cells ( GOLGA4 ), endothelial cells ( DCN, BGN, COL1A2 ), BMP responsive cells ( TTR, RSPO2, LMX1A, MSX1 ), cilium bearing cells ( PCP4, NPHP1 ), oligodendrocytes ( OLIG1, OLIG2 ), astrocytes ( GFAP, AQP4 ) and microglia ( AIF1, CD68 ).

Article Snippet: Levels of FGF8 protein in cell extracts and supernatants were measured using an ELISA kit (Cusabio CSB-E15861h) according to the manufacturer’s instruction.

Techniques: Derivative Assay, Control, Expressing

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Construction, purification, and characterization of FGF21-164. ( A ) FGF21-164 sequence and FGF21 sequence. In the FGF21-164 sequence, red marked the mutated amino acid and blue represents the TR acid sequence from hCD164. In the FGF21 sequence, the red-labeled amino acid represents the amino acid sequence whose n-terminal is deleted in FGF21-164. ( B ) Subcloned cell culture supernatants expressing FGF21-164 and FGF21 were analyzed using Western blotting (Lane M: molecular weight marker. Lane 1~7: FGF21-164 clones 1, 2, 3, A4, B2, F3, and H4. Line 8: FGF21 clone2). ( C ) The purified FGF21-164 and FGF21 were assayed using SDS-PAGE and Western blotting (Lane M: molecular weight marker. Lane 1: purified FGF21-164. Line 2: purified FGF21). ( D ) Purity and hydrodynamic radius of FGF21-164 were analyzed using SEC-HPLC. ( E ) MALDI-TOF mass-spectrometry analysis of FGF21-164. ([M + H] + denotes the singly charged ionic forms, and [M + 2H] 2+ denotes doubly charged ionic forms). ( F ) Circular Dichroism (CD) spectra of FGF21-164 and FGF21.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Purification, Sequencing, Labeling, Cell Culture, Expressing, Western Blot, Molecular Weight, Marker, Clone Assay, SDS Page, Mass Spectrometry, Circular Dichroism

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Glycan forms analysis of the protein FGF21-164. ( A ) O-linked glycan forms of FGF21-164. ( B ) N-linked glycan forms of FGF21-164.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of FGF21-164 in adipocytes induced by 3T3-L1. ( A ) The process of inducing 3T3-L1’s differentiation into adipocytes. ( B ) Reduction in glucose in medium stimulated by FGF21-164 in adipocytes induced by 3T3-L1. Data are means ± SEM (n = 6). * p < 0.05, *** p < 0.001, and **** p < 0.0001 compared to 0 μg/mL FGF21-164. ( C ) Phospho-Erk1/2-specific bands (Thr202/Tyr204; 44–42 kDa) were detected in 3T3-L1-derived adipocytes after FGF21-164 stimulation. ( D ) Evaluation of lipid droplet accumulation in adipocytes induced by 3T3-L1 with or without FGF21-164. The scale bar represents 50 μm.Data are the means ± SEM (n = 6). * p < 0.05, **** p < 0.0001.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Pharmacokinetics of FGF21-164. FGF21-164 plasma levels in C57BL/6 mice as determined via targeted LC-MS after i.v. (tail vein) bolus injection of FGF21-164 protein.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Liquid Chromatography with Mass Spectroscopy, Injection

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: The results of an oral glucose tolerance test applied to ob / ob mice after a single FGF21-164 treatment. ( A ) Blood glucose levels were measured at 0, 2, 3, 4, 5, 6, 7, and 8 h after the administration of FGF21-164. Data are the means ± SEM (n = 4–5). * p < 0.05 and ** p < 0.01 compared with the model. ( B ) The decrease in blood glucose levels post-administration relative to the baseline measurement at 0 h. Data are the means ± SEM (n = 4–5). * p < 0.05 compared with the model.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of a single administration of FGF21-164 on ob / ob mice. ( A , D , G ) Fasting glucose levels. Blood glucose levels were measured at 7, 21, and 28 days after a single dose of FGF21-164 was administered. Data are the means ± SEM (n = 4–5). ns: no significance. * p < 0.05, ** p < 0.01, and **** p < 0.0001. ( B , E , H ) The OGTT at 7, 21, and 28 days. After oral glucose administration, blood glucose levels were measured. Data are the means ± SEM (n = 4–5). * p < 0.05, *** p < 0.001, and **** p < 0.0001 at 6 mg·kg −1 vs. model. # p < 0.05, ## p < 0.01 and #### p < 0.0001 at 12 mg·kg −1 vs. model. ( C , F , I ) The glucose AUC during the OGTT process. Data are the means ± SEM (n = 4–5). ns: no significance. * p < 0.05, *** p < 0.001. ( J ) Body weight measured at 0, 7, 21, and 28 days. Data are the means ± SEM (n = 4–5). ns: no significance.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of repeated administration of FGF21-164. ( A ) Body weight changes. The DIO mice were treated with PBS or FGF21-164 once every 2 days. Data are the means ± SEM (n = 5–7). **** p < 0.0001 vs. Ctrl. #### p < 0.0001 for FGF21-164 vs. the model. ( B ) The liver sections were stained with Oil-red-O. The scale bar represents 100 μm. DIO mice showed many red lipid droplets in hepatic tissue (indicated by the black arrow), while lipid droplet levels were lower after the FGF21-164 treatment.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Staining

Journal: BMJ Open

Article Title: Association between serum vitamin B 6 concentration and risk of osteoporosis in the middle-aged and older people in China: a cross-sectional study

doi: 10.1136/bmjopen-2018-028129

Figure Lengend Snippet: Comparison of serum Vit B 6 and serological metabolism parameters among control, osteopenia and osteoporosis groups

Article Snippet: The concentration of fibroblast growth factor 23 (FGF23) was measured through an ELISA performed using a human FGF23 ELISA kit (CSB-E10113h, Cusabio Biotech, Wuhan, China) and a micro plate reader (MK3, Thermo, Waltham, Massachusetts, USA).

Techniques: Comparison, Control

Journal: BMJ Open

Article Title: Association between serum vitamin B 6 concentration and risk of osteoporosis in the middle-aged and older people in China: a cross-sectional study

doi: 10.1136/bmjopen-2018-028129

Figure Lengend Snippet: Relationship between serum Vit B 6 concentrations and bone metabolism parameter concentrations in control, osteopenia and osteoporosis groups

Article Snippet: The concentration of fibroblast growth factor 23 (FGF23) was measured through an ELISA performed using a human FGF23 ELISA kit (CSB-E10113h, Cusabio Biotech, Wuhan, China) and a micro plate reader (MK3, Thermo, Waltham, Massachusetts, USA).

Techniques: Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 overexpression prevents SCI in mice. (A) Mice were exposed to Ctrl or SCI surgery, and FGF5 mRNA level in the spinal cord was detected at indicating times. (B) FGF5 level was detected using an ELISA kit. (C, D) Mice were intraspinally injected with 2 μL lentivirus carrying FGF5 to overexpress FGF5 in the spinal cord, and FGF5 mRNA and protein levels were detected 2 weeks post‐injection. (E) Mice were overexpressed with FGF5 using lentiviral vectors and then were exposed to SCI or sham surgery 2 weeks post‐injection. BMS score, from 0 (no ankle movement) to 9 (normal gait), was determined at indicating times. (F, G) Sensitivities to mechanical and thermal stimulation were determined 28 days after SCI. (H) Extravasation of Evans blue dye was determined 28 days after SCI. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Over Expression, Enzyme-linked Immunosorbent Assay, Injection, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 knockdown exacerbates SCI in mice. (A, B) Mice were intraspinally injected with 2 μL lentivirus carrying shFGF5 to knockdown FGF5 in the spinal cord, and FGF5 mRNA and protein levels were detected 2 weeks post‐injection. (C) Mice were injected with shFGF5, and then were exposed to SCI or sham surgery 2 weeks post‐injection. BMS score, from 0 (no ankle movement) to 9 (normal gait), was determined at indicating times. (D, E) Sensitivities to mechanical and thermal stimulation were determined 28 days after SCI. (F) Extravasation of Evans blue dye was determined 28 days after SCI. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Knockdown, Injection, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 overexpression reduces inflammation and oxidative stress in SCI mice. (A) Mice were overexpressed with FGF5 using lentiviral vectors, and then were exposed to SCI or sham surgery 2 weeks post‐injection. IL‐6 and TNF‐α levels in the spinal cord were detected. (B) MPO activity in the spinal cord. (C) Western blot images and quantification of p65 phosphorylation. (D) Relative NF‐κB transcription activity. (E, F) Relative levels of NRF2 protein and transcription activity. (G) ROS level detected by DCFH‐DA probe. (H) Quantification of hydrogen peroxide and superoxide anion in spinal cord. (I) The levels of MDA, 3‐NT and 8‐OHdG. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Over Expression, Injection, Activity Assay, Western Blot, Phospho-proteomics, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 knockdown elevates inflammation and oxidative stress in SCI mice. (A) Mice were injected with shFGF5, and then were exposed to SCI or sham surgery 2 weeks post‐injection. IL‐6 and TNF‐α levels in the spinal cord were detected. (B) MPO activity in the spinal cord. (C) Western blot images and quantification of p65 phosphorylation. (D) Relative NF‐κB transcription activity. (E, F) Relative levels of NRF2 protein and transcription activity. (G) ROS level detected by DCFH‐DA probe. (H) Quantification of hydrogen peroxide and superoxide anion in spinal cord. (I) The levels of MDA, 3‐NT and 8‐OHdG. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Knockdown, Injection, Activity Assay, Western Blot, Phospho-proteomics, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 overexpression reduces inflammation, oxidative stress and SCI through activating AMPK. (A) Mice were overexpressed with FGF5 using lentiviral vectors, and then were exposed to SCI or sham surgery 2 weeks post‐injection. Western blot images and quantification of AMPK phosphorylation in the spinal cord were detected. (B) To inhibit AMPK, mice with or without FGF5 overexpression were intraperitoneally injected with CC every 2 days 1 week pre‐SCI. IL‐6 and TNF‐α levels in the spinal cord were detected. (C) ROS level detected by DCFH‐DA probe. (D) The levels of MDA, 3‐NT and 8‐OHdG. (E) BMS score, from 0 (no ankle movement) to 9 (normal gait), was determined at indicating times. (F, G) Sensitivities to mechanical and thermal stimulation were determined 28 days after SCI. (H) Extravasation of Evans blue dye was determined 28 days after SCI. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Over Expression, Injection, Western Blot, Phospho-proteomics, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 overexpression activates AMPK through cAMP/PKA pathway. (A, B) Mice were intraspinally injected with 2 μL lentivirus carrying FGF5 to overexpress FGF5 in the spinal cord, and cAMP level and PKA activity were detected 2 weeks post‐injection. (C) To inhibit PKA, mice with or without FGF5 overexpression were intraperitoneally injected with H89 every 2 days 1 week pre‐SCI. Western blot images and quantification of AMPK phosphorylation in the spinal cord were detected. (D) ROS level detected by DCFH‐DA probe. (E) The levels of MDA, 3‐NT and 8‐OHdG. (F) IL‐6 and TNF‐α levels in the spinal cord were detected. (G) BMS score, from 0 (no ankle movement) to 9 (normal gait), was determined at indicating times. (H, I) Sensitivities to mechanical and thermal stimulation were determined 28 days after SCI. (J) Extravasation of Evans blue dye was determined 28 days after SCI. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Over Expression, Injection, Activity Assay, Western Blot, Phospho-proteomics, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: Serum FGF5 level positively correlates with the sensory and motor function in SCI patients. (A) Serum levels of FGF5 in Ctrl and SCI patients. (B, C) SCI patients were divided into four groups according to serum FGF5 levels, and ASIA sensation and motor scores were measured at indicating groups. n = 56 for Ctrl group and n = 92 for SCI group. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Standard Deviation

Journal: Acta pharmacologica Sinica

Article Title: Transcription factor Klf9 controls bile acid reabsorption and enterohepatic circulation in mice via promoting intestinal Asbt expression.

doi: 10.1038/s41401-021-00850-x

Figure Lengend Snippet: Fig. 2 Klf9 regulation of BA synthesis has a nonhepatic basis. a, b Images of gallbladder and BA levels in the gallbladder, faeces and serum of the control (Klf9flox/flox) and liver-specific Klf9-KO (Klf9alb−/−) mice. c mRNA expression levels of Klf9, Cyp7a1, Cyp8b1, Cyp27a1, Shp, Fxr, Oatp, Mdr2, Bsep, Ntcp and Ost-α in the livers of the Klf9flox/flox and Klf9alb−/−mice. Gene expression was normalised to 36B4 expression. d mRNA expression levels of Klf9, Fgf15, Asbt, Fxr, and Ost-α in the intestines of the Klf9flox/flox and Klf9alb−/−mice. Gene expression was normalised to 36B4 expression. e Protein expression levels of Klf9 and Cyp7a1 in the livers of the Klf9flox/flox and Klf9alb−/−mice. Actin served as the loading control. f Protein expression levels of Klf9, Asbt and Fgf15 in the intestines of the Klf9flox/flox and Klf9alb−/−mice. β-actin served as the loading control. Data are represented as the mean ± SEM. **P < 0.01 (b–d).

Article Snippet: A mouse FGF15 ELISA kit (catalogue: CSB-EL522052MO) was purchased from CUSABIO (Wuhan, China).

Techniques: Control, Expressing, Gene Expression

Journal: Acta pharmacologica Sinica

Article Title: Transcription factor Klf9 controls bile acid reabsorption and enterohepatic circulation in mice via promoting intestinal Asbt expression.

doi: 10.1038/s41401-021-00850-x

Figure Lengend Snippet: Fig. 3 Klf9 affects BA metabolism in the intestine. a Images of gallbladders in the control (Klf9flox/flox) and intestine-specific Klf9-KO (Klf9vil−/−) mice. b BA levels in the gallbladder, serum and faeces of the Klf9flox/flox and Klf9vil−/−mice. c mRNA expression levels of Klf9, Fgf15, Asbt, Fxr and Ost-α in the intestines of the Klf9flox/flox and Klf9vil−/−mice. Gene expression was normalised to 36B4 expression. d mRNA expression levels of Klf9, Cyp7a1, Cyp8b1, Cyp27a1, Shp, Fxr, Oatp, Mdr2, Bsep, Ntcp and Ostα in the liver tissues of the Klf9flox/flox and Klf9vil−/−mice. Gene expression was normalised to 36B4 expression. e Protein expression levels of Klf9 and Cyp7a1 in the intestines of the Klf9flox/flox and Klf9vil−/−mice. β-actin served as the loading control. f Protein expression levels of Klf9, Fgf15, and Asbt in the livers of the Klf9flox/flox and Klf9vil−/−mice. β-actin served as the loading control. Data are represented as the mean ± SEM. *P < 0.05, **P < 0.01 (b–d).

Article Snippet: A mouse FGF15 ELISA kit (catalogue: CSB-EL522052MO) was purchased from CUSABIO (Wuhan, China).

Techniques: Control, Expressing, Gene Expression

Journal: Acta pharmacologica Sinica

Article Title: Transcription factor Klf9 controls bile acid reabsorption and enterohepatic circulation in mice via promoting intestinal Asbt expression.

doi: 10.1038/s41401-021-00850-x

Figure Lengend Snippet: Fig. 4 Klf9 induces Asbt expression in the terminal ileum. Primary small intestinal epithelial cells (a, b) and Caco-2 cells (c, d) were infected with adenovirus with Klf9 overexpression (Klf9) or Klf9 KO (siKlf9). The mRNA expression levels of Klf9, Asbt, and Fgf15 were analysed by RT- qPCR. Gene expression was normalised to 36B4 expression. Dual-luciferase reporter gene assays of the (e) Fgf15 promoter (–2500 bp) and (f) ABST promoter (−2500 bp) were performed in HEK-293A cells. g The wild-type putative Klf9-binding element and its mutant sequence in the Asbt promoter region. A series of Asbt promoters fused to the luciferase reporter gene (–2500, –1850, –700, and –150 bp) were cotransfected into HEK-293A cells together with Klf9 expression plasmids or pc-DNA3.1 (control, Ctl). h ChIP assays were performed to confirm Klf9 binding to the Asbt gene promoter. Data are represented as the mean ± SEM. *P < 0.05 (a–f).

Article Snippet: A mouse FGF15 ELISA kit (catalogue: CSB-EL522052MO) was purchased from CUSABIO (Wuhan, China).

Techniques: Expressing, Infection, Over Expression, Quantitative RT-PCR, Gene Expression, Luciferase, Binding Assay, Mutagenesis, Sequencing, Control

Journal: Acta pharmacologica Sinica

Article Title: Transcription factor Klf9 controls bile acid reabsorption and enterohepatic circulation in mice via promoting intestinal Asbt expression.

doi: 10.1038/s41401-021-00850-x

Figure Lengend Snippet: Fig. 5 Klf9 overexpression promotes BA reabsorption. a Images of gallbladders in the control (Rosa) and intestine-specific Klf9- overexpressing (Klf9Rosa+/+) mice. b BA levels in the gallbladder, serum and faeces of the Rosa and Klf9Rosa+/+ mice. c mRNA expression levels of Klf9, Fgf15, Asbt, Fxr and Ost-α in the intestines of the Rosa and Klf9Rosa+/+ mice. Gene expression was normalised to 36B4 expression. d mRNA expression levels of Klf9, Cyp7a1, Cyp8b1, Cyp27a1, Shp, Fxr, Oatp, Mdr2, Bsep, Ntcp and Ost-α in the livers of the Rosa and Klf9Rosa+/+

Article Snippet: A mouse FGF15 ELISA kit (catalogue: CSB-EL522052MO) was purchased from CUSABIO (Wuhan, China).

Techniques: Over Expression, Control, Expressing, Gene Expression

Journal: Acta pharmacologica Sinica

Article Title: Transcription factor Klf9 controls bile acid reabsorption and enterohepatic circulation in mice via promoting intestinal Asbt expression.

doi: 10.1038/s41401-021-00850-x

Figure Lengend Snippet: Fig. 6 Changes in small intestine morphology. a HE staining of the jejunum, ileum and colon in flox/flox mice, Klf9vil−/−mice, Rosa mice and Klf9Rosa+/+ mice; images are shown at ×200 magnification; scale bars, 100 μm. b Fgf15 protein concentration in blood from the four groups. c Immunohistochemical staining of Asbt in the ileums of the four groups; images are shown at ×400 magnification; scale bars, 50 μm. d Representative immunofluorescence staining of Fgf15 in the intestinal tissues in the four groups; images are shown at ×400 magnification; scale bars, 50 μm. *P < 0.05.

Article Snippet: A mouse FGF15 ELISA kit (catalogue: CSB-EL522052MO) was purchased from CUSABIO (Wuhan, China).

Techniques: Staining, Protein Concentration, Immunohistochemical staining

Journal: Acta pharmacologica Sinica

Article Title: Transcription factor Klf9 controls bile acid reabsorption and enterohepatic circulation in mice via promoting intestinal Asbt expression.

doi: 10.1038/s41401-021-00850-x

Figure Lengend Snippet: Fig. 7 Proposed model of Klf9 induction of BAs absorption. Proposed model of Klf9 induction of BA absorption. Klf9 promotes Asbt expression in the ileum, and Asbt promotes BA reabsorption. The reabsorbed BAs activate the Fxr/Fgf15 signalling pathway. Fgf15 is secreted from the intestine to repress Cyp7a1 expression in the liver and reduce BA production.

Article Snippet: A mouse FGF15 ELISA kit (catalogue: CSB-EL522052MO) was purchased from CUSABIO (Wuhan, China).

Techniques: Expressing

Journal: Redox Biology

Article Title: Exercise and dietary intervention ameliorate high-fat diet-induced NAFLD and liver aging by inducing lipophagy

doi: 10.1016/j.redox.2020.101635

Figure Lengend Snippet: Exercise increased the FGF21 levels in serum and muscle. (A) FGF21 level in serum by ELISA. (B) FGF21 protein levels in liver and muscle by Western Blot. (C) FGF21 content in liver and muscle by ELISA. (D) FGF21 mRNA levels in liver and muscle by qPCR. Data are shown as the means ± SEM, n = 8 per group. Significance was designated by asterisks with *p < 0.05.

Article Snippet: Rat FGF21 enzyme-linked immunosorbent assay (ELISA) kit was obtained from Cusabio Biotech (Cat. No. CSB-EL008627RA, Cusabio Biotech, Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Redox Biology

Article Title: Exercise and dietary intervention ameliorate high-fat diet-induced NAFLD and liver aging by inducing lipophagy

doi: 10.1016/j.redox.2020.101635

Figure Lengend Snippet: FGF21 attenuated LDs accumulation in FFA-treated HepG2 cells through an autophagy dependent way. HepG2 cells were treated with FFA (400 μM). (A) Representative confocal images of LDs by BODIPY 493/503 staining. Scale: 50 μm. (B) TG level in HepG2 cells. (C) Representative immunoblotting of PLIN2, LC3II/LC3I and p62 in HepG2 cells. HepG2 cells were pre-transfected with Atg5 shRNA or empty vector by lenti-virus, then treated with FFA (400 μM) with or without FGF21 (1 μg/ml). (D) Representative confocal images of LDs by BODIPY 493/503 staining. Scale: 50 μm. (E) TG level in HepG2 cells. Experiments were repeated three times. Data are shown as the means ± SEM. Significance was designated by asterisks with *p < 0.05.

Article Snippet: Rat FGF21 enzyme-linked immunosorbent assay (ELISA) kit was obtained from Cusabio Biotech (Cat. No. CSB-EL008627RA, Cusabio Biotech, Wuhan, China).

Techniques: Staining, Western Blot, Transfection, shRNA, Plasmid Preparation, Virus

Journal: Redox Biology

Article Title: Exercise and dietary intervention ameliorate high-fat diet-induced NAFLD and liver aging by inducing lipophagy

doi: 10.1016/j.redox.2020.101635

Figure Lengend Snippet: FGF21 increased lipophagy through AMPK-dependent pathway in FFA-treated HepG2 cells. HepG2 cells were treated with FFA (400 μM), FGF21 (1 μg/ml) as well as AMPK inhibitor-Dorsomorphin (1 μmol/L). (A) Representative of p -AMPK (Thr172)/AMPK total, p -ULK1 (Ser555)/ULK1 total and LC3-II/LC3-I immunoblotting in HepG2 cells treated with FFA, FGF21 and Dorsomorphin, and the quantified data. (B) Representative confocal images of HepG2 cells expressing GFP-RFP-LC3 and quantitation of early autophagosome puncta and autolysosome puncta following FFA, FGF21 and Dorsomorphin treatment. Yellow showed co-localization of GFP and RFP, indicating early autophagosomes. Red only showed autolysosomes, Scale: 20 μm. Experiments were repeated three times. Data are shown as the means ± SEM. Significance was designated by asterisks with *p < 0.05. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Rat FGF21 enzyme-linked immunosorbent assay (ELISA) kit was obtained from Cusabio Biotech (Cat. No. CSB-EL008627RA, Cusabio Biotech, Wuhan, China).

Techniques: Western Blot, Expressing, Quantitation Assay

Journal: Redox Biology

Article Title: Exercise and dietary intervention ameliorate high-fat diet-induced NAFLD and liver aging by inducing lipophagy

doi: 10.1016/j.redox.2020.101635

Figure Lengend Snippet: FGF21 increased the lipophagy and reduced the lipid accumulation in FFA-treated HepG2 cells. HepG2 cells were treated with FFA (400 μM), FGF21 (1 μg/ml) as well as AMPK inhibitor-Dorsomorphin (1 μmol/L). (A) Representative confocal images of colocalization of LDs and autophagosomes by BODIPY 493/503 staining in GFP-LC3 protein expressed HepG2 cells. Scale: 50 μm. (B) TG level in HepG2 cells treated with FFA, FGF21 and Dorsomorphin. Experiments were repeated three times. Data are shown as the means ± SEM. Significance was designated by asterisks with *p < 0.05.

Article Snippet: Rat FGF21 enzyme-linked immunosorbent assay (ELISA) kit was obtained from Cusabio Biotech (Cat. No. CSB-EL008627RA, Cusabio Biotech, Wuhan, China).

Techniques: Staining

Journal: Redox Biology

Article Title: Exercise and dietary intervention ameliorate high-fat diet-induced NAFLD and liver aging by inducing lipophagy

doi: 10.1016/j.redox.2020.101635

Figure Lengend Snippet: FGF21 ameliorated the cell senescence through AMPK dependent autophagic pathway in FFA-treated WI-38 cells. WI-38 cells were treated with FFA (400 μM), FGF21 (1 μg/ml) as well as AMPK inhibitor-Dorsomorphin (1 μmol/L). (A) Representative of p -AMPK (Thr172)/AMPK total, p -ULK1 (Ser555)/ULK1 total and LC3-II/LC3-I immunoblotting in WI-38 cells and quantified data. (B) TG level in WI-38 cells. (C) Representative SA-β-gal staining (Scale: 50 μm), (D) SA-β-gal activity, (E) lipofuscin and (F) MDA contents in WI-38 cells. (G) p16 and (H) p27 mRNA levels in WI-38 cells. Experiments were repeated three times. Data are shown as the means ± SEM. Significance was designated by asterisks with *p < 0.05.

Article Snippet: Rat FGF21 enzyme-linked immunosorbent assay (ELISA) kit was obtained from Cusabio Biotech (Cat. No. CSB-EL008627RA, Cusabio Biotech, Wuhan, China).

Techniques: Western Blot, Staining, Activity Assay