Journal: Cell communication and signaling : CCS

Article Title: FGF19 increases mitochondrial biogenesis and fusion in chondrocytes via the AMPKα-p38/MAPK pathway.

doi: 10.1186/s12964-023-01069-5

Figure Lengend Snippet: Fig. 1 FGF19 induces a transient increase in mitochondrial number and an enhanced generation of ATP products. a Representative TEM images showing the changes of mitochondrial number in chondrocytes induced by FGF19 at 200 ng/ml in the presence of KLB (200 ng/ml). The images were chosen based on three independent experiments (n = 3). Orange arrows indicated individual mitochondrion. b Quantification of mitochondrial number (per cell) in chondrocytes induced by FGF19 at 200 ng/ml in the presence of KLB (200 ng/ml). Quantitative analyses of the mitochondrial number were based on nine cells (per group) from three independent experiments (n = 3). c Representative immunofluorescent staining showing the number changes of mitochondria in living chondrocytes induced by FGF19 (200 ng/ml) in the presence of KLB (200 ng/ml) for 72 h. The images were chosen based on three independent experiments (n = 3). Red, individual mitochondrion; Green, F-actin; Blue, nucleus. d Linear quantification of fluorescence intensity of mitochondrion number in chondrocytes induced by FGF19 at 200 ng/ml in the presence of KLB (200 ng/ml) by Image Pro Plus 6.0. e ATP assay showing the increase of intracellular ATP products in chondrocytes induced by FGF19 at 200 ng/ml in the presence of KLB (200 ng/ml). The results were based on three independent experiments (n = 3). f Representative western blotting showing the expression change of CS in chondrocytes induced by FGF19 at 200 ng/ml in the presence of KLB (200 ng/ml). The images were chosen based on three independent experiments (n = 3). g Quantification of CS by western blotting in (f). The data in b are shown as box (from 25, 50 to 75%) and whisker (minimum to maximum values) plots. The significant difference analysis in b, e and g was based on Student T-test

Article Snippet: FGF19 (200 ng/ml, No.100-32, PEPRO TECH, USA) and/or KLB (200 ng/ ml, 2619-KB-050, R&D Systems, USA) at a 1:1 ratio were added into the culture media as the experimental group and continued to incubate for 72 h. Thaw all the components of Cell NavigatorTM Mitochondrion Staining Kit at RT before starting the experiment.

Techniques: Staining, Fluorescence, ATP Assay, Western Blot, Expressing, Whisker Assay

Journal: Cell communication and signaling : CCS

Article Title: FGF19 increases mitochondrial biogenesis and fusion in chondrocytes via the AMPKα-p38/MAPK pathway.

doi: 10.1186/s12964-023-01069-5

Figure Lengend Snippet: Fig. 4 FGF19 activates p38/MAPK signalling in chondrocytes. a RNA sequencing showing the changes in the expression of MAPK-related mediators in chondrocytes induced by FGF19 at 200 ng/ml in the presence of KLB (200 ng/ml). Three pairs of samples were obtained from three independent cell isolates (n = 3), namely, samples 1, 1′, 1′′and 1′′′, samples 2, 2′, 2′′and 2′′′, and samples 3, 3′, 3′′and 3′′′. The data were present as log2(FPKM + 1). b Representative western blotting showing the expression change of ERK, p-ERK, p38, p-p38, JNK and p-JNK in chondrocytes induced by FGF19 at 200 ng/ml in the presence of KLB (200 ng/ml). The images were chosen based on three independent experiments (n = 3). c Quantification of p38 and p-p38 by western blotting in (b). d Representative immunofluorescent staining showing the change in the expression and distribution of p-p38 in chondrocytes induced by FGF19 (200 ng/ml) in the presence of KLB (200 ng/ml) for 72 h. The images were chosen based on three independent experiments (n = 3). Red, p-p38; Green, F-actin; Blue, nucleus. e Quantification of fluorescence intensity of p-p38 in chondrocytes induced by FGF19 at 200 ng/ml in the presence of KLB (200 ng/ml). The data were based on nine cells from three independent experiments. The data in e were shown as box (from 25, 50 to 75%) and whisker (minimum to maximum values) plots. The significant difference analysis in c and e was based on Student T-test

Article Snippet: FGF19 (200 ng/ml, No.100-32, PEPRO TECH, USA) and/or KLB (200 ng/ ml, 2619-KB-050, R&D Systems, USA) at a 1:1 ratio were added into the culture media as the experimental group and continued to incubate for 72 h. Thaw all the components of Cell NavigatorTM Mitochondrion Staining Kit at RT before starting the experiment.

Techniques: RNA Sequencing, Expressing, Western Blot, Staining, Fluorescence, Whisker Assay

Journal: Cell communication and signaling : CCS

Article Title: FGF19 increases mitochondrial biogenesis and fusion in chondrocytes via the AMPKα-p38/MAPK pathway.

doi: 10.1186/s12964-023-01069-5

Figure Lengend Snippet: Fig. 5 Inhibition of p38 attenuated FGF19-enhanced AMPKα activity. a Representative western blotting showing the expression change of p38, p-p38, AMPKα, p-AMPKα, PGC-1α and SIRT1 in chondrocytes induced by SB203580 (10 µM) in the presence of FGF19 (200 ng/ml) and KLB (200 ng/ml). The images were chosen based on three independent experiments (n = 3). b Quantification of p38, p-p38, AMPKα, p-AMPKα, PGC-1α and SIRT1 by western blotting in (a). c Representative immunofluorescent staining showing the change in the distribution of p-AMPKα in chondrocytes induced by SB203580 (10 µM) in the presence of FGF19 (200 ng/ml) and KLB (200 ng/ml). The images were chosen based on three independent experiments (n = 3). Red, p-AMPKα; Green, F-actin; Blue, nucleus. d Representative immunofluorescent staining showing the change in the expression and distribution of PGC-1α in chondrocytes induced by SB203580 (10 µM) in the presence of FGF19 (200 ng/ml) and KLB (200 ng/ml). The images were chosen based on three independent experiments (n = 3). Red, PGC-1α; Green, F-actin; Blue, nucleus. e Quantification of fluorescence intensity of p-AMPKα and PGC-1α in chondrocytes induced by SB203580 (10 µM) in the presence of FGF19 (200 ng/ml) and KLB (200 ng/ml). The data in e were shown as box (from 25, 50 to 75%) and whisker (minimum to maximum values) plots. The significant difference analysis in b and e was based on Student T-test

Article Snippet: FGF19 (200 ng/ml, No.100-32, PEPRO TECH, USA) and/or KLB (200 ng/ ml, 2619-KB-050, R&D Systems, USA) at a 1:1 ratio were added into the culture media as the experimental group and continued to incubate for 72 h. Thaw all the components of Cell NavigatorTM Mitochondrion Staining Kit at RT before starting the experiment.

Techniques: Inhibition, Activity Assay, Western Blot, Expressing, Staining, Fluorescence, Whisker Assay