Figure 5. R406 can upregulate cellular H₂S level and H₂S can promote chemical reprogramming.

• A
  H₂S levels in DMSO‐ and R406‐treated reprogramming intermediates detected by monobromobimane method. n = 4.
• B
  H₂S levels in reprogramming intermediates treated with DPBS, HA, and NAC. n = 3.
• C
  Immunofluorescence of Sall4 in reprogramming intermediates treated by R406 + HA and R406 + DPBS on d12. n = 3. Scale bar, 100 μm.
• D
  RT–qPCR analysis of Sall4, Cdh1, Epcam, and Esrrb gene expression in DPBS‐ and HA‐treated cells on d12. n = 3.
• E
  Immunofluorescence of Sall4 in NAC‐ and DPBS‐treated cells on d12. n = 5. Scale bar, 100 μm.
• F
  RT–qPCR analysis of Sall4, Cdh1, Epcam, and Esrrb gene expression in DPBS‐ and NAC‐treated cells on d12. n = 3.
• G
  H₂S levels in reprogramming intermediates treated with shNC and shCbs. n = 3.
• H
  Diagram showing the procedure of reprogramming infected with shRNA for Cbs.
• I
  RT–qPCR analysis of Cbs expression in MEFs infected with shNC and shCbs. n = 3.
• J, K
  Immunofluorescence of Sall4 in shNC‐ and shCbs‐infected cells with and without R406 treatment on d12. n = 5. Scale bar, 100 μm.
• L
  Diagram showing the procedure of reprogramming infected with Cbs‐OE (pSin‐Cbs) virus.
• M
  RT–qPCR analysis of Cbs expression in MEFs infected with pSin‐GFP^N and pSin‐Cbs viruses. n = 3.
• N
  Immunofluorescence of Sall4 in WT cells and Cbs overexpressing cells on d12. n = 4. Scale bar, 100 μm.

Data information: All data are presented as mean ± SD. Statistical significance was assessed by the two‐tailed Student’s t‐test, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. See also Fig EV5.