1. Decreased expression of SAM68 in human testes with spermatogenic defects 
Le-Jun Li, M.D., Feng-Bin Zhang, M.D., Shu-Yuan Liu, M.D., Yong-Hong Tian, M.D., Fang Le, M.D., Hang-Ying Lou, M.D., He-Feng Huang, M.D., Fan Jin, M.D. 
Fertility and Sterility 
Volume 102, Issue 1, Pages e3 (July 2014)
DOI: /j.fertnstert
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Relative testicular RNA transcript expression of SAM68. Gene expression was normalized to the expression of the housekeeping gene GAPDH. *P<.0001.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Representative gels for SAM68 protein levels in the testes of infertile patients. GAPDH was used as a loading control.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Immunohistochemical analysis of SAM68 expression in the testes of infertile patients. (A) Testicular tissue section of a representative OAZ patient with normal spermatogenesis. (B) Testicular tissue section of a representative patient with Sertoli cell–only syndrome. (C, D) Testicular tissue section of two representative patients with MA. Moderate or strong staining for SAM68 protein. (E, F) Testicular tissue section of two representative patients with MA. Negative or weak staining for SAM68 protein. Relevant cells are indicated by arrows. SPC = spermatocytes; MS = mature spermatids; SC = Sertoli cells. Final magnification ×400.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
Effect of Sam68 on the proliferation and apoptosis rate of GC-2spd(ts) cells. (A) Cell proliferation was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Data are presented as the mean of two experiments with five replicates samples. Error bars indicate the 95% confidence intervals. Statistically significant increases in cell proliferation are indicated by the asterisks (P<.05). (B) The apoptosis rate was analyzed using Annexin V/propidium iodide staining. Transfected Sam68 shRNA vector induced significant apoptosis (P<.05; n = 3). Two independent stable clones are shown, referred to as GC2-Sam68-G3 and GC2-Sam68-F2.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

6. Supplemental Figure 1
GC-2spd(ts) cells depleted of endogenous Sam68. GC-2spd(ts) cells transfected with an empty vector (pGPU6/Hygro-SHNC) or a vector containing a shRNA (Sam68 shRNA) were selected with hygromycin B, and knockdown populations depleted of Sam68 were identified. Two independent stable clones are shown, as indicated. (A) RT-PCR was carried out on total cellular RNA. The DNA fragments were visualized on agarose gels stained with ethidium bromide. The expression of Sam68 was examined as well as the expression of controls including GAPDH and ACTB. (B) Reduction in Sam68 protein was analyzed by immunoblotting with anti-Sam68 antibody and anti-GAPDH antibodies as loading controls.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions