Fig 1: CD147 promotes sperm motility and induces the acrosome reaction(A) CASA results showing the effect of recombinant CD147 (rCD147; 10 µg/mL) and CD147-neutralizing antibody (anti-CD147 Ab, 10 µg/mL) on sperm motility. The same amount of vehicle or normal IgG was used as a control. (B) CASA results showing the effect of rCD147 (10 µg/mL) and progesterone (P4, 20 µM) on the hyperactivated motility of capacitated sperm. (C) FITC-PSA staining results showing the effect of rCD147 (10 µg/mL), P4 (20 µM), the ionophore A23187 (5 µM), or the indicated combined treatments on the sperm acrosome reaction. (D) FITC-PSA staining results showing the effect of conditioned medium of CD147 knockdown (shCD147/CM) or control (shNC/CM) KGN cells on the acrosome reaction in the presence or absence of a CD147-neutralizing antibody (10 µg/mL). (E and F) Hyaluronan binding assay (E) and human sperm-hamster oocyte penetration assay (F) results showing the effect of rCD147 (10 µg/mL) and CD147-neutralizing antibody (anti-CD147 Ab, 10 µg/mL) on the hyaluronan binding ability and oocyte-penetrating ability of treated sperm. The same amount of vehicle or normal IgG was used as a control. Data are presented as the mean ± SEM.
Fig 2: CD147 deficiency and impairment of the acrosome reaction in the sperm of patients with asthenozoospermia(A) Representative confocal images showing the expression and localization of CD147 (green) in the head and midpiece before capacitation in normal sperm. The intensity of CD147 on the sperm head was increased after capacitation (red arrowheads). Nuclei were counterstained with DAPI (blue). Scale bar, 10 µm. (B) Quantification of fluorescent signals showing a significant increase in the integrated density of CD147 in the normal sperm head region after capacitation. (C) Representative western blot of normozoospermic (normal; n = 26) and asthenozoospermic patient samples (Astheno; n = 21) showing decreased CD147 expression in asthenozoospermic patients compared with normal men. GAPDH was used as a loading control. (D) Quantification of CD147 expression in sperm from the normal and asthenozoospermia cohorts. (E) qPCR assay of CD147 mRNA levels in sperm from the normal (n = 28) and asthenozoospermia groups (n = 29). (F) Computer-assisted sperm analysis of the motility of sperm from normozoospermic men (normal, n = 26) and asthenozoospermic patients (Astheno, n = 21). (G) Quantification of the ionophore-induced (A23187) acrosome reaction measured by FITC-PSA staining. The acrosome reaction of sperm from patients with asthenozoospermia (Astheno, n = 7) was significantly lower than that from normozoospermic men (normal, n = 10). Data are presented as the mean ± SEM.
Fig 3: CD147 induces Ca2+ influx in human sperm(A) Representative fluorescence images of human sperm loaded with Fluo-4, a Ca2+-sensitive dye, before (basal) and after (rCD147) treatment with rCD147 (10 µg/mL) in Ca2+-free or Ca2+-containing media with or without CD147-neutralizing antibody (10 µg/mL). Sperm samples treated with P4 (500 nM) were used as a positive control. (B) Representative time course changes in Fluo-4 intensity (F) normalized to the initial intensity (F0) in the human sperm treated with rCD147 or P4 as in (A). (C) Quantification of changes in intracellular Ca2+ in sperm samples treated as in (A). The calcium values of responsive sperm were calculated. Numbers in brackets represent the number of sperm recorded in at least three independent experiments. (D) Quantification of rCD147-induced (10 µg/mL) changes in intracellular Ca2+ in sperm samples of normal men under noncapacitated and capacitated conditions. Data are presented as the mean ± SEM.
Fig 4: Soluble CD147 interacts with sperm-bound CD147(A) ELISA results showing the presence of soluble CD147 in human follicular fluid (n = 15). Sample dilution buffer in the ELISA kit was used as a blank control. (B) Western blot results showing the expression of CD147 in vector control-transfected (shNC) or CD147 shRNA-transfected (shCD147) KGN cells. ß-actin was used as a loading control. (C) ELISA results showing the presence of soluble CD147 in serum-free conditioned medium obtained from CD147 knockdown (shCD147) or control (shNC) KGN cells. (D) PLA results showing the interactions between His-tagged CD147 and membrane-bound CD147 on human sperm with or without capacitation. Human sperm that endogenously expressed membrane-bound CD147 were treated with His-tagged rCD147, which lacks the C-terminal cytoplasmic tail. The potential interaction was detected by PLA using anti-His and anti-CD147 (C terminus) antibodies (red signals in bottom panel). An enlarged image is shown on the bottom right panel. Sperm samples labeled with either PLA only (top left), anti-His antibody with PLA (top middle), or anti-CD147 antibody with PLA (C terminus) (top right) were used as controls. Scale bar, 10 µm. Data are presented as the mean ± SEM.
Fig 5: Recombinant CD147 treatment improves functions in infertile sperm(A) CASA results showing the effect of a CD147-neutralizing antibody (10 µg/mL) and/or rCD147 treatment (10 µg/mL) in sperm obtained from patients with asthenozoospermia. (B) FITC-PSA staining results showing the effect of rCD147 (10 µg/mL) on the acrosome reaction of sperm obtained from patients with asthenozoospermia in the presence or absence of the CD147-neutralizing antibody (10 µg/mL). (C) Hyaluronan binding assay results showing the effect of the CD147-neutralizing antibody (anti-CD147 Ab, 10 µg/mL) and/or rCD147 (10 µg/mL) on the hyaluronan binding ability of sperm obtained from patients with asthenozoospermia. (D) Correlation analysis of seminal plasma CD147 levels as determined by ELISAs with the fertilization rate in IVF procedures. (E) ELISA results showing the level of CD147 in the seminal plasma of infertile men with the indicated IVF outcome. Data are presented as the mean ± SEM.
Supplier Page from Abcam for Human BSG ELISA Kit (CD147)