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. 2020 Mar 11;9(6):e010240. doi: 10.1161/JAHA.118.010240

Figure 2.

Figure 2

mPinX1t is a protein encoding gene. A and B, High‐density sucrose gradient polysome fractionation was performed using mESC lysate. A, Nondenaturing RNA agarose gel showing the presence and the integrity of 28S and 18S rRNAs in different fractions. Fractions 1 to 4 represent free ribonucleoprotein fractions; fractions 5 to 8 represent 40S, 60S fractions; fractions 9 to 11 represent monosome fractions; fractions 12 to 22 represent polysome fractions. B, PCR reactions of different fractions showing the presence of mPinX1t transcripts in monosome and polysome fractions. Equal volume of PCR product was loaded onto each lane. “–ve” represents negative control. C, Representative western blot showing the expressions of mPinX1 and mPinX1t proteins in undifferentiated mESCs and mESC differentiation derivatives at day 7+5. β‐Tubulin was used as the loading control. mPinX1 was observed at around 45 kDa and mPinX1t was observed between 15 and 25 kDa. D, Representative western blot showing the expression of mPinX1 proteins in undifferentiated mESCs using an antibody targeting the C‐terminal of mPinX1 (Orb47163). mPinX1 was observed at around 45 kDa; mPinX1t, which lacks the C‐terminal of mPinX1, was not detected. mESC indicates mouse embryonic stem cell; RNPs, ribonucleoproteins; PCR, polymerase chain reaction.