Fig. 4: Okinawa rail, Japanese ptarmigan, and Blakiston’s fish owl-derived iPSC growth in knockout serum replacement (KSR) and low molecular inhibitor medium.

a, b Cell growth analysis of Okinawa rail, Japanese ptarmigan, and Blakiston’s fish owl-derived iPSCs in serum base medium and knockout serum replacement (KSR) medium. Images show the morphology of the iPSC colonies after alkaline phosphatase staining (a). The serum base medium (i–vi) and the KSR base medium (vii–xii) represent. Okinawa rails (i, iv, vii, and x), Japanese ptarmigans (ii, v, viii, and xi), and Blakiston’s fish owls (iii, vi, ix, and xii) shows. The bars represent 500 μm. Cell numbers of Okinawa rail, Japanese ptarmigan, and Blakiston’s fish owl iPSCs in serum-based and KSR-based media (b). Yellow bars represent Okinawa rail, green bars represent Japanese ptarmigans, and purple bars represent Blakiston’s fish owl. Centerlines of box plots indicate medians; box limits indicate the 25th and 75th percentiles. n = 6. **P < 0.01. c Alkaline phosphatase staining of Okinawa rail, Japanese ptarmigan, and Blakiston’s fish owl-derived iPSCs in medium containing JAK inhibitor. Those panels show the control medium (i–vi), medium containing 1 μM JAK inhibitor (vii–xii), and medium containing 10 μM JAK inhibitor (xiii–xviii). Okinawa rail (i, iv, vii, x, xiii, and xvi), Japanese ptarmigans (ii, v, viii, xi, xiv, and xvii), and Blakiston’s fish owl (iii, vi, ix, xii, xv, and xviii) -derived iPSCs shows. The bars represent 500 μm. d Counting of Okinawa rail, Japanese ptarmigan, and Blakiston’s fish owl-derived iPSCs in medium containing JAK inhibitor. Yellow bars show Okinawa rail, green bars show Japanese ptarmigans, and purple bars show Blakiston’s fish owl-derived iPSCs. Centerlines of box plots indicate medians; box limits indicate the 25th and 75th percentiles. n = 6. *P < 0.05. e Alkaline phosphatase staining of Okinawa rail, Japanese ptarmigan, and Blakiston’s fish owl-derived iPSCs in medium containing FGFR inhibitor. Those panels show the control medium (i–vi), medium containing 1 μM JAK inhibitor (vii–xii), and medium containing 10 μM JAK inhibitor (xiii–xviii). Okinawa rail (i, iv, vii, x, xiii, and xvi), Japanese ptarmigans (ii, v, viii, xi, xiv, and xvii), and Blakiston’s fish owl (iii, vi, ix, xii, xv, and xviii) -derived iPSCs shows. The bars represent 500 μm. f Counting of Okinawa rail, Japanese ptarmigan, and Blakiston’s fish owl-derived iPSCs in medium containing FGFR inhibitor. Centerlines of box plots indicate medians; box limits indicate the 25th and 75th percentiles. n = 6. *P < 0.05. g Cellular morphology of Okinawa rail, Japanese ptarmigan, and Blakiston’s fish owl-derived iPSCs in medium containing Go6983 (PKC inhibitor) and medium lacking Go6983. Bright-field images (i–vi) and alkaline phosphatase (AP) staining images (vii–xii) show. The medium containing Go6983 additive (i–iii and vii–ix) and the medium lacking Go6983 minus (iv–vi and x–xii) show. The Okinawa rail (i, iv, vii, and x), Japanese ptarmigans (ii, v, viii, and xi), and Blakiston’s fish owl (iii, vi, ix, and xii) shows. The bars represent 500 μm. h Summary of serum and signal dependency.